Chauhan, O. P - Non-Thermal Processing of Foods-Chapman and Hall - CRC (2019)

Non-thermal Processing of Foods
Non-thermal Processing
of Foods
Edited by
O. P. Chauhan
CRC Press
Taylor & Francis Group
6000 Broken Sound Parkway NW, Suite 300
Boca Raton, FL 33487-2742
© 2019 by Taylor & Francis Group, LLC

CRC Press is an imprint of Taylor & Francis Group, an Informa business
No claim to original U.S. Government works
Printed on acid-free paper
International Standard Book Number-13: 978-1-1380-3584-3 (Hardback)
This book contains information obtained from authentic and highly regarded sources. Reasonable efforts have been made
to publish reliable data and information, but the author and publisher cannot assume responsibility for the validity of all
materials or the consequences of their use. The authors and publishers have attempted to trace the copyright holders of all
material reproduced in this publication and apologize to copyright holders if permission to publish in this form has not been
obtained. If any copyright material has not been acknowledged please write and let us know so we may rectify in any future
reprint.
Except as permitted under U.S. Copyright Law, no part of this book may be reprinted, reproduced, transmitted, or utilized
in any form by any electronic, mechanical, or other means, now known or hereafter invented, including photocopying,
microfilming, and recording, or in any information storage or retrieval system, without written permission from the
publishers.
For permission to photocopy or use material electronically from this work, please access www.copyright.com (http://
www.copyright.com/) or contact the Copyright Clearance Center, Inc. (CCC), 222 Rosewood Drive, Danvers, MA 01923,
978-750-8400. CCC is a not-for-profit organization that provides licenses and registration for a variety of users. For
organizations that have been granted a photocopy license by the CCC, a separate system of payment has been arranged.
Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are used only for
identification and explanation without intent to infringe.
Library of Congress Cataloging‑in‑Publication Data
Names: Chauhan, O. P., author.

Title: Non-thermal processing of foods / O.P. Chauhan.
Description: Boca Raton : Taylor & Francis, a CRC title, part of the Taylor &
Francis imprint, a member of the Taylor & Francis Group, the academic
division of T&F Informa, plc, 2019. | Includes bibliographical references.
Identifiers: LCCN 2018033439 | ISBN 9781138035843 (hardback : acid-free paper)
Subjects: LCSH: Food--Preservation.
Classification: LCC TP371.2 .C44 2019 | DDC 664/.028--dc23
LC record available at https://lccn.loc.gov/2018033439
Visit the Taylor & Francis Web site at

http://www.taylorandfrancis.com
and the CRC Press Web site at

http://www.crcpress.com
Contents
Editor ................................................................................................................................................ix
Contributors ......................................................................................................................................xi
Chapter 1
High-Pressure Processing: Principles and Engineering Aspects ...................................................... 1
Simran Kaur Arora and O. P. Chauhan
Chapter 2
High Hydrostatic Pressure Processing of Cereals and Pulses ......................................................... 11
Sajad Ahmad Wani and Pradyuman Kumar
Chapter 3
Effect of High-Pressure Processing on Selected Food Processing Operations ............................... 27
Jincy M. George and Navin Kumar Rastogi
Chapter 4
High-Pressure Processing of Meat, Fish, and Poultry Products ...................................................... 47
K. Jayathilakan, Khudsia Sultana and M. C. Pandey
Chapter 5
High-Pressure Processing of Milk and Milk Products .................................................................... 69
Ashish Kumar Singh, Sanket Borad, G. S. Meena, Heena Sharma, and Sumit Arora
Chapter 6
Pulsed Electric Field Processing: Principles and Engineering Aspects .......................................... 89
R. Singh, B. P. Kaur, and S. Thangalakshmi
Chapter 7
PEF Processing of Fruits, Vegetables, and their Products ............................................................. 107
R. Kumar, S. Vijayalakshmi, T. Kathiravan, and S. Nadanasabapathi
Chapter 8
Pulse Electric Field Processing of Milk and Milk Products ......................................................... 129
Neelam Upadhyay, C. T. Manoj Kumar, Heena Sharma, Sanket Borad,
and Ashish Kumar Singh
Chapter 9
Application of Ultrasonic in Food Processing ............................................................................... 145
Pradeep Singh Negi and Navin Kumar Rastogi
v
vi Contents
Chapter 10
Use of Pulsed Light in Food Processing ........................................................................................ 173
Lakshmi E. Unni and O. P. Chauhan
Chapter 11
Ozone Application in Food Processing ......................................................................................... 189
C. T. Manoj Kumar and Latha Sabikhi
Chapter 12
Improving the Efficacy of Ozone Treatment in Food Preservation ............................................... 213
Shyam Ramkrishna Garud, Pradeep Singh Negi, and Navin Kumar Rastogi
Chapter 13
High-Pressure CO2 Processing of Foods ....................................................................................... 235
Bindvi Arora, Alka Joshi, and Shruti Sethi
Chapter 14
Pulsed Magnetic Field Processing of Foods .................................................................................. 261
K. Chitravathi and O. P. Chauhan
Chapter 15
Use of Plasma in Food Processing ................................................................................................. 283
A. K. Pandey and O. P. Chauhan
Chapter 16
Electron Beam Processing of Foods .............................................................................................. 315
Shima Shayanfar and Suresh D. Pillai
Chapter 17
Combination of Non-thermal Processes and their Hurdle Effect .................................................. 329
Swati Sethi, Rahul Kumar Anurag, Yogesh Kumar, and O. P. Chauhan
Chapter 18
Non-thermal Processing of Seafoods ............................................................................................ 373
K. Sarika and J. Bindu
Chapter 19
Packaging Requirements for Non-thermal Processed Foods ........................................................ 395
Poonam Mishra and O. P. Chauhan
Contents vii
Chapter 20
Commercialization and Regulatory Issues of Non-thermal Processed Foods .............................. 417
Feby Luckose, B. S. Mamatha, and O. P. Chauhan
Chapter 21
Consumer Acceptance and Future Trends of Non-thermal-Processed Foods ............................... 433
Prerna Nath, S. J. Kale, and Bharat Bhushan
Index .............................................................................................................................................. 455
Editor
Dr. O. P. Chauhan is with the Defence Research Development Services (DRDS) under the Defence
Research and Development Organization (DRDO), Ministry of Defence, Government of India. He
is currently working as Scientist and Head of the Fruits and Vegetables Technology and Material
Management Group at the Defence Food Research Laboratory, Mysore, India. He is also Secretary
of the Specialist Panel on Food Science, Hill Agriculture, Biofuels and Bioresources of Life Sciences
Research Board of DRDO. Dr. Chauhan earned an MSc and a PhD in Food Technology from G.
B. Pant University of Agriculture and Technology, Pantnagar, India. His research interests include
postharvest handling of fruits and vegetables including high pressure processing, pulsed electric
field processing, microwave dehydration and modified/controlled atmosphere packaging/storage,
among others. His research findings have appeared in more than 75 international and national peer-
reviewed journals. He has edited one book and published nine international and six national book
chapters. He also has nine patents to his credit. He has presented 30 oral and 60 poster papers in
various national and international conferences. He is the chief editor of American Journal of Food
Technology and editorial board member of Journal of Food Science and Technology, International
Journal of Food and Fermentation Technology, among others. Dr. Chauhan has supervised
four PhD and several MSc, MTech, and BTech students. He is a recipient of the Indian National
Science Academy (INSA) fellowship to work at the German Institute of Food Technologies (DIL),
Quakenbrück, Germany, on Advanced Food Technologies, besides being trained on high pressure
technology in the United Kingdom. He has been instrumental in organizing several conferences,
workshops, training programs, and courses. He has transferred 22 technologies to 36 firms for
commercialization.
Dr. Chauhan is a recipient of the DRDO Young Scientist Award, AFSTI Young Scientist
Award, Laboratory Scientist of the Year Award, Laljee Godhoo Smarak Nidhi Award (AFSTI),
DRDO Technology Group Award, FICCI Best Postharvest Technology Innovation Award, DRDO
Technology Spin-off Award, Bioved Fellowship, Dr. J. S. Pruthi Award (AFSTI & AIFPA), as well
as several Best Paper & Poster Awards from different associations. He is a Life Member of the
Association of Food Scientists and Technologists (India), Nutrition Society of India and the Indian
Science Congress.
ix
Contributors
Rahul Kumar Anurag Shyam Ramkrishna Garud
Agricultural Structures and Environmental Department of Food Engineering
Control Division Central Food Technological Research Institute,
ICAR-Central Institute of Post Harvest Mysore, India
Engineering and Technology
Ludhiana, India Jincy M. George
Academy of Scientific and Innovative Research
Bindvi Arora and Non-thermal Processing Laboratory
Division of Food Science and Postharvest Department of Food Engineering
Technology Central Food Technological Research Institute
ICAR-Indian Agricultural Research Institute Mysore, India
New Delhi, India
K. Jayathilakan
Freeze Drying and Animal Products
Simran Kaur Arora
Technology Division
Department of Food Science and Technology
Defence Food Research Laboratory
G.B. Pant University of Agriculture and
Mysore, India
Technology
Pantnagar, India Alka Joshi
Division of Food Science and Postharvest
Sumit Arora Technology
ICAR-National Dairy Research Institute ICAR-Indian Agricultural Research Institute
Karnal, India New Delhi, India
Bharat Bhushan S. J. Kale

Horticultural Crop Processing Division Horticultural Crop Processing Division
ICAR-Central Institute of Post Harvest ICAR-Central Institute of Post Harvest
Engineering and Technology Engineering and Technology
Abohar, India Abohar, India
T. Kathiravan
J. Bindu
Computational Modeling and Nanoscale
Fish Processing Division
Processing Unit
ICAR-Central Institute of Fisheries Technology
Indian Institute of Food Processing Technology
Cochin, India
Thanjavur, India
Sanket Borad B. P. Kaur
Dairy Technology Division Department of Food Engineering
ICAR-National Dairy Research Institute National Institute of Food Technology
Karnal, India Entrepreneurship and Management
Sonepat, India
O. P. Chauhan
Defence Food Research Laboratory Pradyuman Kumar
Mysore, India Department of Food Engineering and
Technology
K. Chitravathi Sant Longowal Institute of Engineering and
Defence Food Research Laboratory Technology
Mysore, India Longowal, India
xi
xii Contributors
R. Kumar Prerna Nath

Food Engineering and Packaging Technology Horticultural Crop Processing Division
Division ICAR-Central Institute of Post Harvest
Defence Food Research Laboratory Engineering and Technology
Mysore, India Abohar, India
Yogesh Kumar
Pradeep Singh Negi
Agricultural Structures and Environmental
Department Fruit and Vegetable Technology
Control Division
CSIR-Central Food Technological Research
ICAR-Central Institute of Post Harvest
Institute
Engineering and Technology
Mysore, India
Ludhiana, India
Feby Luckose A. K. Pandey

PG Department of Food Science and Defence Food Research Laboratory
Technology Mysore, India
St. Aloysius College (Autonomous)
Mangalore, India M. C. Pandey
Freeze Drying and Animal Products
B. S. Mamatha
Technology Division
Department of Biological Sciences
Nitte University Center for Science Education
Mysore, India
and Research (NUCSER)
Mangalore, India
Suresh D. Pillai
C. T. Manoj Kumar National Center for Electron Beam Research
Dairy Technology Division Texas A&M University
ICAR-National Dairy Research Institute College Station, Texas
Karnal, India
Navin Kumar Rastogi
G. S. Meena
Academy of Scientific and Innovative Research
ICAR- National Dairy Research Institute
and Non-thermal Processing Laboratory
Karnal, India
Department of Food Engineering
Poonam Mishra Central Food Technological Research
Department of Food Engineering and Institute
Technology Mysore, India
ICAR- National Dairy Research Institute
Karnal, India Latha Sabikhi
Tezpur University Dairy Technology Division
Tezpur, India ICAR-National Dairy Research Institute
Karnal, India
S. Nadanasabapathi
Food Engineering and Packaging Technology K. Sarika
Division Fish Processing Division
Defence Food Research Laboratory ICAR-Central Institute of Fisheries Technology
Mysore, India Cochin, India
Contributors xiii
Shruti Sethi Khudsia Sultana

Division of Food Science and Postharvest Freeze Drying and Animal Products
Technology Technology Division
ICAR-Indian Agricultural Research Institute ICAR- National Dairy Research Institute
New Delhi, India Karnal, India
Mysore, India
Swati Sethi
Foodgrains and Oilseeds Processing Division S. Thangalakshmi
ICAR-Central Institute of Post Harvest National Institute of Food Technology
Engineering and Technology Entrepreneurship and Management
Ludhiana, India Sonepat, India
Lakshmi E. Unni
Heena Sharma Fruits and Vegetables Technology Division
Dairy Technology Division Defence Food Research Laboratory
ICAR-National Dairy Research Institute Mysore, India
Karnal, India
Neelam Upadhyay
Dairy Technology Division
Shima Shayanfar
ICAR-National Dairy Research Institute
National Center for Electron Beam Research
Karnal, India
Texas A&M University
College Station, Texas S. Vijayalakshmi
Computational Modeling and Nanoscale
R. Singh Processing Unit
National Institute of Food Technology Indian Institute of Food Processing Technology
Entrepreneurship and Management Thanjavur, India
Sonepat, India
Sajad Ahmad Wani
Department of Food Engineering and
Ashish Kumar Singh Technology
Dairy Technology Division Sant Longowal Institute of Engineering and
ICAR-National Dairy Research Institute Technology
Karnal, India Longowal, India
ChaptEr 1
high-pressure processing
Principles and Engineering Aspects
Simran Kaur Arora and O. P. Chauhan
CONtENtS
1.1 HPP Technology ....................................................................................................................... 1

1.1.1 Principles Governing the Effects of HPP on Food ....................................................... 2
1.1.2 Global Manufacturers of HPP Units .............................................................................4
1.2 High Pressure Equipment and Engineering Aspects................................................................ 4
1.3 Conclusion ................................................................................................................................7
References ..........................................................................................................................................7
There is an increasing demand for fresh and natural food products free from preservatives and
chemical additives throughout the world. With the advancements and improved understanding
about the process of high-pressure processing (HPP) and its effect on the quality and shelf-life of
the product, there has been noted a global growth in the application of this technology at commer-
cial scale for the manufacture of products. One of the most attractive features of HPP is that the
HPP processed foods have high resemblance to the unprocessed counterparts in terms of nutritional
and sensory properties. This unique characteristic and less dependence on added flavours, colour,
and acid/salt/sugar and class II preservatives (e.g., sulphite, nitrite, benzoate, sorbate) make HPP
superior to the traditional thermal processes and was selected as one of the best innovations in food
processing (Dunne, 2005). There is an increasing trend among consumers for consuming more
natural and fresh foods. According to a report by Nielsen (2015), 57% of respondents from Asia-
Pacific, Europe, Middle East/Africa, Latin America, and North America consume more natural and
fresh foods while more than 40% of respondents say the absence of artificial colours, flavours, and
foods made from vegetables/fruits are very important. In this regard, HPP promises processing of
foods in a healthier manner with the goodness of natural foods but with enhanced shelf-life.
1.1 hpp tEChNOLOGY
HPP is also known as High Hydrostatic Pressure Processing, Ultrahigh-Pressure Processing,

and Pascalization. HPP employs application of high hydrostatic pressure to the food in an enclosed
vessel for a certain period of time sufficient to inactivate harmful pathogenic and vegetative spoil-
age microorganisms and selective enzymes. The applied pressure may range from 100 to 1000 MPa
1
2 non-tHerMAL ProCessinG oF FooDs
(whereas the normal atmospheric pressure is equal to 0.1 MPa). The process temperature may also
vary over a wide range of below 0°C to above 100°C. The lower temperatures may be employed
where the product is sensitive to heat damage while the higher temperatures are needed to destroy
the microbial spores. For example, to attain a significant reduction in the Clostridium sporogenes
spore count, high pressure of 690 MPa at 80°C for 20 min was found to be effective (Crawford
et al., 1996). However, Clostridium botulinum and its spores, which are the most heat-resistant
and most lethal to human beings, are the most pressure resistant and could pose a threat to safety.
Such microorganisms which adapt themselves to high-pressure conditions are known as Piezophiles
(Saldo, 2016) and the process of application of heat along with high pressure in order to kill micro-
bial spores is also known as pressure-assisted thermal process (PATP). According to Martínez
and Balasubramaniam (2016) FDA had issued letters of no objection to two industrial petitions for
preserving shelf-stable low-acid samples by PATP. Though the sensory and nutritional quality of
food is maintained best if HPP is employed at low temperature (0°C–40°C), regulatory bodies and
governments are concerned more with the microbiological safety of food products. Presence of
moisture (40% or more) in food products is also essential for anti-microbial effect of HPP (Muntean
et al., 2016). HPP not only preserves food from harmful microorganisms and undesirable enzymes
but also offers protection to functional compounds like antioxidants (Matser et al., 2004), probi-
otics (Penna et al., 2007; Cruz et al., 2010), colostrum, and lactoferrin and some such preserva-
tion techniques have been patented by certain food giants, for example Fonterra, New Zealand
(Huppertz, 2010). Interestingly, content of some functional compounds like gamma-aminobutyric
acid in brown rice, buck wheat, and soybean; vitamin C in potato and avocado; and lycopene in
tomato has been found to increase by HPP (Ohara et al., 2015). This is attributed to “high-pressure
induced transformation” (also known as Hi-Pit) (Yamazaki, 2006). This is making HPP technology
very attractive and promising to the food researchers to look out for the possibilities of developing
new products with enhanced functional attributes.
1.1.1 principles Governing the Effects of hpp on Food
There are basically four principles which govern the effect of HPP on the quality of food.
1. Le Chatelier’s principle: It states that any reaction, conformational change, or phase transition
which is accompanied by decrease in volume is enhanced under high-pressure conditions (Yordanov
and Angelova, 2010) and the processes which involve volume increase are inhibited by pressure
(Butz and Tauscher, 1998). Therefore, at a relatively low temperature (0°C–40°C) covalent bonds
are unaffected by HPP whereas hydrophobic and ionic interactions responsible for the tertiary and
quaternary structures of molecules are altered at more than 200 MPa (Hendrickx et al., 1998).
Therefore, HPP, unlike thermal processing, neither harms ascorbic acid (Oley et al., 2006), folates
(Butz et al., 2004), anthocyanins (Verbeyst et al., 2010), lycopene (Gupta et al., 2010), conjugated
linoleic acid (Martinez-Monteagudo and Saldana, 2014), or essential amino acids nor causes toxic
compounds to develop (Damodaran, 1996). HPP changes inter-atomic distances, thus affecting
those interactions for which bonding energy depends on distance (Martinez-Monteagudo et al.,
2012). However, for PATP, based on Gibbs’s definition of free energy, change in temperature dur-
ing pressure treatment causes change in volume and energy. Reactions such as phase transitions or
molecular reorientation depend on both temperature and pressure (Balasubramaniam et al., 2015).
Also, in PATP the process time is significantly reduced (3–15 min) from the process dependent on
thermal or pressure treatment alone.
2. Isostatic pressing: It is also known as Pascal’s Principle. It states that in HPP, food is compressed
and uncompressed uniformly from all the directions irrespective of the size and shape of the food
product, unlike in thermal processing (Yaldagard et al., 2008; Huppertz, 2010). This protects the
food from getting damaged and deshaped. Proteins are flexible and compressible. Globular proteins
HiGH-Pressure ProCessinG 3
get denatured with the application of high pressure due to the presence of some void spaces in the
interior of their molecular structure leading to compressibility. The average partial specific volume,
given by vº, of globular proteins in the hydrated state is about 0.74 mL/g. According to Gonzalez and
Tanchuco (1977) the partial specific volume is the sum of three components:
v o = VC + VCav + ∆VSol
where VC is the sum of the atomic volumes, VCav is the sum of the volumes of the void spaces in
the interior of the protein, and ∆VSol is the volume change due to hydration (Gekko and Hasegawa,
1986). The larger the vº of a protein, the larger is the contribution of void spaces to partial spe-
cific volume, and the more unstable the protein will be when pressurized. Fibrous proteins are
mostly devoid of void spaces, and hence, they are more stable to hydrostatic pressure than globular
proteins. Pressure-induced denaturation of globular proteins is usually accompanied by a reduction
in volume of about 30–100 mL/mol mainly because of the elimination of void spaces as the protein
unfolds, and hydration of the nonpolar amino acid residues that become exposed during unfold-
ing. Pressure-induced protein denaturation at low pressure is highly reversible. In the case of
pressure-denatured oligomeric proteins and enzymes, subunits first dissociate at 0.001–2 kbar, and
then subunits denature at higher pressures (Weber, 1992); removal of pressure results in subunit
reassociation and almost complete restoration of enzyme activity after several hours. High hydro-
static pressures (2–10 kbar) irreversibly damage cell membranes and cause dissociation of organelles
in microorganisms, inactivating vegetative microorganisms. Hydrostatic pressure of 1–7 kbar for
30 min at 25°C causes gelation of egg white, 16% soy protein solution, or 3% actomyosin solution
(Okomoto et al., 1990). Hydrostatic pressure of 1–3 kbar causes meat tenderization due to partial
fragmentation of muscle myofibrils (Suzuki et al., 1990). However, a pressure indicator developed
using copper tablet challenges the principle of isostatic pressing when placed in the geometric centre
of a large ham, indicating that the ham received approximately 9 MPa less pressure than the HHP
system delivered (Minerich and Labuza, 2003).
3. Microscopic ordering principle: It states that at a constant temperature, an increase in pressure
increases the degree of ordering of molecules of a given substance. So, at higher pressure, molecules
will be less disordered and such chemical reactions will take place in which the resulting molecules
will have higher degree of ordering. However, nature prefers a high degree of disorder. Therefore,
during HPP free energy (entropy) of the system is increased. It further increases during PATP and
molecular arrangement will depend upon the temperature and pressure applied along with the change
in volume. Normally, increase in temperature increases volume and degree of disorder of molecules,
while increase in pressure causes increase in the order of molecules and decrease in volume.
4. Arrhenius relationship: The various reaction rates are influenced by thermal effects during HPP.
The net effects can be synergistic, additive, or antagonistic (Balasubramaniam et al., 2015). When
pressure build-up is rapid in the HPP system and there are minimal chances of energy getting dis-
sipated to the surroundings, compressive or adiabatic heating of the pressure transmitting fluid is
observed. For water it is 2°C–3°C/100 MPa and for fats and oils it is 4°C–9°C/100 MPa (Huppertz,
2010). In a perfectly insulated (adiabatic) system, the product should return to its initial temperature
upon decompression. In practice, however, the product returns to a temperature slightly lower than
its initial temperature as a result of some heat losses to the environment during the compression
phase. The increased temperature of the medium and thus product (both due to heat transfer from
the medium and self-heating due to the presence of moisture and fat in the food itself) may help
achieving the desired high temperature during PATP to kill spores. High pressure (400–600 MPa)
and high temperature (>70°C) treatment have a synergistic effect on spore inactivation (Patazca
et al., 2006; Reddy et al., 2006; Ahn et al., 2007; Bull et al., 2009; Daryaei et al., 2013). When the
intensity of treatment is above a certain threshold (400–600 MPa and 90°C–120°C), the treatment
is often referred to as PATP, pressure-assisted thermal sterilization, or ultra-high-pressure thermal
sterilization. Strains of Clostridium botulinum are the target pathogens in some PATP treatment
studies (Reddy et al., 2013; Skinner et al., 2014).
1.1.2 Global Manufacturers of hpp Units
The Japanese were first in exploring HPP, in setting up HPP equipment manufacturing units, and
in launching HPP-processed food products commercially. Later, research took place in almost all the
developed countries and today the major equipment manufacturers are mainly located in America,
Europe, and Asian countries. The various scientists in these manufacturing companies, along with
the other scientific fraternity, loaned this equipment to run their trials, researched various aspects of
HPP on food, its constituents and shelf-life. Below is the list of some manufacturers who help set up
HPP units on the lab/pilot/commercial scale. Some of them also have option for lending HPP units
for processing products on a charge basis to other parties. This kind of arrangement helps such food
producers who are interested in employing HPP to preserve their food products/innovate new food
products without investing in the purchase of HPP units, and in turn reduces the cost of production
for the owners who own high-cost HPP units.
1. USA: Avure Technologies, Engineered Pressure Systems Incorporated, Harwood Engineering,

Elmhurst Research
2. Japan: Kobe Steel, Toyo Koatsu Co. Ltd., Mitsubishi Heavy Industries Ltd
3. China: Wenzhou Binyi Machinery Co. Ltd., BaoTou KeFa High Pressure Technology Co. Ltd
4. Spain: Hiperbaric
5. France: ACB Pressure System-Alstom
6. Poland: Unipress
7. Germany: Multivac, UHDE Hochdrucktechnik
8. Netherlands: Resato Internationals
9. UK: Stansted Fluid Power
10. Sweden: Flow International Corporation
Details about some specific models produced by the above manufacturers have been informatively
compiled by Balasubramaniam et al. (2016) and Berg et al. (2001).
1.2 hIGh prESSUrE EQUIpMENt aND ENGINEErING aSpECtS
The HPP equipment typically consists of:
1. A pressure vessel: This is the most important part of HPP in which the sample is placed either in
the pre-packaged form or as such in the case of liquid food items. In the former case, a pressure
transmitting fluid is needed, which may be water, some water-oil emulsion, food grade water-glycol
solutions, silicone oil, sodium benzoate solutions, ethanol solutions, or castor oil (Balasubramaniam
et al., 2015). One may decide the type of pressure transmitting fluid on the basis of its corrosion-
resistance properties, fluid viscosity changes under pressure, ability to seal under pressure, and heat
of compression. The inner side of the inside cylinder, which is in direct contact with water or food,
should be corrosion resistant and is preferably made up of stainless steel (Vetter et al., 2001) while
those made up of tough material like high-tensile steel (low alloy) may require addition of approved
anticorrosive agents in the pressure transmitting fluid. The pressure vessel is cylindrical and may be
placed in horizontal, vertical, or tilting position (for ease in loading and unloading process). In the
horizontal position it is easy and fast to load and unload the food material while in the vertical position
loading and unloading are difficult, time consuming, and may require the help of a crane, but the pres-
sure transmitting fluid may be reused (Berg et al., 2001). The volume of the vessel may range from
0.1 to 2 L (for the laboratory use), 10–25 L (pilot plant), and 100 L and above (Batch/Commercial
production). The pressure vessel may be one piece (monobloc vessel) or may comprise two or more
concentric cylinders. Monobloc vessels operate under pressure less than 400 MPa and have diameters
less than 15 cm. In modern technology of two or more concentric cylinders, the outer cylinders press
the inner cylinders such that the wall of the pressure chamber is always under some residual stress.
According to the Safety Codes given by ASME (Section 8, Division 3 of the Boiler and Pressure
Vessel Code), the inner cylinder should “leak before break” so as to relieve the pressure in a con-
trolled manner and prevent any burst and accident. Normally the thickness of the vessel is decided
primarily by the maximum pressure the system is required to maintain. However, the outer cylinder
may be wire wound around an inner cylinder (wire winding increases strength-to-weight ratio and
improves the fatigue life [Sedighi and Jabbari, 2013]; the cylinder inner diameter shrinks because of
the pressure from the wire leading to compressive residual stresses inside the vessel wall) or encapsu-
lated in a liquid-filled layer or autofrettage or heat shrink to produce prestressed vessel in which there
is permanent pressurization of the outer cylinder, thus enabling pressure to reach 680 MPa or more.
In the autofrettage process, first, a very high pressure is applied to the cylinder by either dragging a
mandrel through the shell or by pumping oil into it, resulting in plastic deformation of the innermost
bore and the lesser stressed outside expands elastically. Then the pressure is released and the elasti-
cally deformed outside of the shell tries to regain its original shape but is prevented by the plastically
deformed inside part, causing permanent stress in the shell (Son et al., 2012; Koutchma, 2014). In the
heat-shrink technique, the external cylinder is heated until it expands, and the inner cylinder is cooled
until it shrinks. Then, the outer layer is assembled over the inner layer (shrink fitting). On cooling the
whole assembly to room temperature, residual hoop stresses develop, putting the inner cylinder in
compression (Moss and Basic, 2013).
2. Two end closures: At the two ends of the cylindrical vessel, closures are used to cover it after loading
and on the external surface of the closures, pressure is applied.
3. Yoke: Yoke is a structure used for restraining end closures while under high axial pressure. The yoke
may comprise a number of steel plates as used in the equipment manufactured by ACB Pressure
Systems-Alstom Hyperbar or a wire-wound steel frame as present in Flow Quintus System.
4. A pressure-creating device: A low-pressure pump and an intensifier are normally used to gener-
ate desired pressure in the system. The low-pressure pump is used for creating direct, piston-type
compression whereas the high-pressure intensifier is needed for indirect compression. Generally, the
former is present in laboratory or pilot plant units to have fast compression while the latter is used
to create very high pressures in commercial scale units. Compression time is a function of pump
horsepower. For example, it takes 3–4 min for a 100-horsepower pump to bring a 50-L vessel to
an operating pressure of 680 MPa. Degree of compression required to reach the desired pressure
depends on the compressibility of the processing fluid and product. Compressibility of water (the
most commonly used medium) is 10% and 15% at 300 MPa and 600 MPa, respectively (Huppertz,
2010). On compression, water gets reduced in volume, so additional water is needed to achieve the
high pressure. Sometimes where HPP is performed under high-temperature conditions, the heating
of the pressure transmitting fluid is performed, which expands and helps build the extra pressure.
For best pressure transmission, the ideal food for HPP has no gas inclusions and no empty spaces
in the package (Muntean et al., 2016). Air is removed from the vessel before the HPP treatment to
reduce pumping costs by eliminating air compression. However, residual air has no effect on micro-
bial inactivation kinetics of HPP treated, packaged food. Uniform temperature increase and control
is needed for the successful application of this technology (Farkas and Hoover, 2000).
5. Instrumentation and controls: For the proper operation of HPP plant, one must have controls for:
a. Pressure measurement: Metallic membrane cell (for pressure up to 400 bar), Boudon-gauge
type (for pressure up to 7000 bar), and strain gauge transducers (for pressure up to 15,000 bar)
may be employed (Spain and Paauwe, 1977). Vertical plunger in a close-fitting cylinder with
calibrated weights is used for the calibration of pressure measuring instruments.
b. Temperature measurement: In HPP either thermocouples (250°C–1000°C) or resistant
thermometers (230°C–2800°C) are used for measuring temperature changes during the
process.
c. Flow measurement: Various devices are available for measuring flow depending on the
viscosity of the fluid and the low- or high-pressure side of the process. Under high pressure
up to 4000-bar turbine flow meter is used while for flow measurement under 400-bar Coriolis
flow meter with small flow rates is used.
d. Level measurement: For the continuous system of HPP it is necessary to know the level of
product in the vessel for continuous and safe running of the equipment. Several systems are
available to measure it under high pressure. Sight glass systems, level float sensors, and
membrane- or gas-based systems may be used depending on the type of food under process
and the process conditions.
HPP Cycle: HPP operates on electricity and does not create any waste product (Yaldagard et al.,
2008). In a typical batch-type HPP cycle, the food sample may or may not be packaged and is placed
in the vessel, which is already partially filled with processing fluid. The vessel is subsequently
closed and then the system is pressurized to the desired pressure (normally at the rate of 5 MPa
per sec). The sample is maintained at that pressure for the required holding time, which may range
from a few seconds up to several hours. However, the holding time in general up to 10 min have
commercial relevance. Following the holding time, pressure is released (normally it takes less than
30 s in a commercial unit) and the vessel can be unloaded. In total the five major steps of loading,
compression, holding time, decompression, and unloading take as much time in batch operation as
in batch process retorting for thermal sterilization. To reduce the total operation time several devel-
opments in the design of HPP equipment have occurred. For example, horizontal vessel position is
preferred over vertical to eliminate the requirement for a hoist or crane to load the product into the
vessel and for possible semi-automation for loading and unloading of the vessel. Also, two or more
vessels may be used in parallel to increase the throughput as it ensures that at least one vessel is at
pressure all times while the other vessel is either under loading or unloading process. This kind of
arrangement leads to semi-continuous operation. One intensifier may be used for two or more pres-
sure vessels used in such arrangements. In certain cases, particularly for dairy operations, the vessel
volume has been increased up to 600 L with processing time up to 15 min in order to handle large
amount of liquid milk in shorter time than before (Huppertz, 2010). Microbial efficacy of the pulsed
HPP process has been reported to be more effective than an equivalent single pulse of equal time.
In pulsed HPP, product is subjected to compression-decompression cycles of fixed pressure holding
time. However, rapid compression and decompression increase the number of cycles on the vessel
and subjects the vessel material to enormous stress and reduces its life.
Development of high-pressure microscopy had enabled the visualization of the motility of
molecular motors under conditions of high-pressure such as kinesin, F1-ATPase, and bacterial
flagellar motors (Nishiyama, 2015) and it was reported that applied pressure works directly to
weaken the intermolecular interaction between tubulin molecules (an important component of the
cellular cytoskeleton).
Flexible packaging material that can compress by about 15% with no structural damage is
normally used for packaging food products before HPP; examples include, polyester, polypropylene,
and polyethylene. Delamination, changes in the polymer structure, sealing properties, and oxygen
transmission rate are some factors that need to be considered while selecting packaging material
for HPP treatment particularly at high temperatures. Vacuum-packed products in flexible packaging
are suggested as most appropriate for HPP treatment (Muntean et al., 2016). However, environmen-
tally friendly, degradable packaging material is the demand of modern consumers to protect the
environment. Therefore, future efforts should focus on testing the suitability of biodegradable and
environmentally safe packaging material for HPP technology.
Currently, investment costs of high-pressure equipment may range from 0.6 to 4 million USD
depending upon various choices (Balasubramaniam et al., 2016). Generally, vessel closures and
yoke account for 50%–60% of the capital cost, while pumping system accounts for 30%–35% and
process control and allied instrumentation accounts for 10%–15%. The HPP-processed foods need
refrigeration after processing for transportation and storage till usage. The HPP technique is expen-
sive and thus the products are also costlier than their thermally processed counterparts. Nonetheless,
in one of the consumer reports, more than 90% of respondents in Latin America, Asia-Pacific, and
Africa/Middle East and close to 80% of respondents in Europe and North America showed their
willingness to pay premium for foods with health attributes (Nielsen’s Survey, 2015). This gives
motivation to continue pursuing research and development in the HPP technique, process,
equipment instrumentation, and automation.
1.3 CONCLUSION
In comparison to thermal processing, HPP has an edge in retaining more nutrients and flavour
compounds. It is being developed largely to cater to those consumers who demand foods that are
nutritionally healthier, more natural, and minimally processed with no preservatives. HPP is gov-
erned by Le Chatelier’s Principle for Chemical Reactions, the Principle of Microscopic Ordering,
Isostatic Pressing, and the Arrhenius Relationship. HPP may be achieved in batch process or
semi-continuous mode. Over the years there have been various advancements in the design and
understanding of HPP technology, equipment, and accessories. Though its popularity is growing
worldwide, the high cost of production and food safety when operated at low temperatures remain
concerns. A major breakthrough is needed to make HPP affordable by the people of developing
countries.
rEFErENCES
Ahn, J., Balasubramaniam, V. M., and Yousef, A. E. (2007). Inactivation kinetics of selected aerobic and
anaerobic bacterial spores by pressure-assisted thermal processing. Int. J. Food Microbiol. 113:321–329.
Balasubramaniam, V. M., Canovas, G. V. B., and Lelieveld, H. L. M. (2016). High-pressure processing equip-
ment for the food industry. In High Pressure Processing of Food, pp. 39–65. Springer Science + Business
Media, New York. doi:10.1007/978-1-4939-3234-4_3.
Balasubramaniam, V. M., Monteagudo, S. I. M., and Gupta, R. (2015). Principles and application of high-
pressure based technologies in the food industry. Annu. Rev. Food Sci. Technol. 6:435–462.
Berg, R. W. V. D., Hoogland, H., Lelieveld, H. L. M., and Schepdael, L. V. (2001). High pressure equipment
designs for food processing applications. In Ultra High Pressure Treatments of Foods (Eds.) M. E. G.
Hendrickx, D. Knorr, L. Ludikhuyze, A. V. Loey and V. Heinz, pp. 297–313. Springer Science + Business
Media, Boston, MA.
Bull, M. K., Oliver, S. A., Diepenbeek, R. J., Kormelink, F., and Chapman, B. (2009). Synergistic inactivation
of spores of proteolytic Clostridium botulinum strains by high pressure and heat is strain and product
dependent. Appl. Environ. Microbiol. 75:434–445.
Butz, P., Serfert, Y., Fernandez-Garcia, A., Dieterich, S., Lindauer, R. et al. (2004). Influence of high-pressure
treatment at 25 degrees C and 80 degrees C on folates in orange juice and model media. J. Food Sci.
69:S117–S121.
Butz, P., and Tauscher, B. (1998). Food chemistry under high hydrostatic pressure. High Pressure Food Sci,
Biosci and Chem. 133–144.
Crawford, Y. J., Murano, E. A., Olson, D. G., and Shenoy, K. (1996). Use of high pressure and irradiation to
eliminate Clostridium sporogenes spores in chicken breast. J. Food Prot. 59:711–715.
Cruz, A. G. D., Faria, J. D. A. F., Saad, S. M. I., Bolini, H. M. A., Ana, A. S. S., and Cristianini, M. (2010).
High pressure processing and pulsed electric fields: Potential use in probiotic dairy foods processing.
Trends Food Sci. Technol. 21:483–493.
Damodaran, S. (1996). Amino acids, peptides, and proteins. Food Chemistry, 3rd edn. (Ed.) O. R. Fennema,
pp. 362–439. Marcel Dekker, New York.
Daryaei, H., Balasubramaniam, V. M., and Legan, J. D. (2013). Kinetics of Bacillus cereus spore inactivation
in cooked rice by combined pressure-heat treatment. J. Food Protect. 76:616–623.
Dunne, C. P. (2005). High pressure keeps food fresher. SSC-Natick Press Release #05-22, May 3. http://www.
natick.army.mil/about/pao/05/05-22.htm.
Farkas, D. F., and Hoover, D. G. (2000). High pressure processing. J. Food Sci. 64(4):47–63.
Gekko, K., and Y. Hasegawa. (1986). Compressibility-structure relationship of globular proteins. Biochemistry
25:6563–6571.
Gonzalez, O. N., and Tanchuco, R. H. (1977). Chemical composition and functional properties of coconut
protein isolate. Phil. J. Coco. Studies. 11:21–29.
Gupta, R., Balasubramaniam, V. M., Schwartz, S. J., and Francis, D. M. (2010). Storage stability of lycopene
in tomato juice subjected to combined pressure-heat treatments. J. Agric. Food Chem. 58:8305–8313.
Hendrickx, M., Ludikhuyze, L., Van den Broeck, I., and Weemaes, C. (1998). Effects of high pressure on
enzymes related to food quality. Trends Food Sci Technol. 9:197–203.
Huppertz, T. (2010). High pressure processing of milk. In Improving the Safety and Quality of Milk. Vol. 1
Milk production and processing. (Ed.) M. W. Griffiths, pp. 373–399. Woodhead Publishing, New Delhi,
India.
Koutchma, T. (2014). Adapting High Hydrostatic Pressure (HPP) for Food Processing Operations. Academic
Press Elsevier, London, UK.
Martinez-Monteagudo, S. I., Saldana M. D. A., Torres J. A., and Kennelly J. J. (2012). Effect of pressure-
assisted thermal sterilization on conjugated linoleic acid (CLA) content in CLA-enriched milk. Innov.
Food Sci. Emerg.Technol. 16:291–297.
Martinez-Monteagudo, S. I., and Saldana, M. D. A. (2014). Modeling the retention kinetics of conjugated
linoleic acid during high-pressure sterilization of milk. Food Res. Int. 62:169–176.
Martínez-Monteagudo, S. I., and Balasubramaniam, V. M. (2016). Fundamentals and applications of high-
pressure processing technology. In High Pressure Processing of Foods (Eds.) V. M. Balasubramaniam,
G. V. Barbosa-Cánovas and H. L. M. Lelieveld, pp. 3–17. Springer, New York.
Matser, A. M., Krebbers, B., van den Berg, R. W., and Bartels, P. V. (2004). Advantages of high pressure ster-
ilization on quality of food products. Trends Food Sci. Technol. 15:79–85.
Minerich, P. L., and Labuza, T. P. (2003). Development of a pressure indicator for high hydrostatic pressure
processing of foods. Innov. Food Sci. Emerg. Technol. 4:235–243.
Moss, D. R., and Basic, M. (2013). High pressure vessels. In Pressure Vessel Design Manual (4th ed.) (Eds.)
D. R. Moss and M. Basic, pp. 473–556. Butterworth-Heinemann, Oxford, UK.
Muntean, M. V., Marian, O., Barbieru V., Catunescu, G. M., Ranta, O., Drocas, I., and Terhes, S. (2016).
High pressure processing in food industry–characteristics and applications. Agric. Agric. Sci. Procedia.
10:377–383.
Nielsen (2015). Nielsen global health and wellness report-January 2015.pdf. We are what we eat-healthy eating
trends around the world. www.nielsen.com.
Nishiyama, M. (2015). High-pressure microscopy for studying molecular motors. In: High Pressure Bioscience
(Subcellular Biochemistry, Vol. 72). (Eds.) K. Akasaka and H. Matsuki, pp. 593–612. Springer
Science+Business Media, Dordrecht, the Netherlands. doi:10.1007/978-94-017-9918-8_27.
Ohara, E., Kawamura, M., Ogino, M., Hoshino, E., Kobayashi, A. Hoshino, J., Yamazaki, A., and Nishiumi, T.
(2015). Application of high-pressure treatment to enhancement of functional components in agricultural
products and development of sterilized foods. In High Pressure Bioscience (Subcellular Biochemistry,
Vol. 72). (Eds.) K. Akasaka and H. Matsuki, pp. 567–589. Springer Science+Business Media, Dordrecht,
the Netherlands. doi:10.1007/978-94-017-9918-8_26.
Okomoto, M., Kawamura, Y., and Hayashi, R. (1990). Application of high pressure to food processing: Textural
comparison of pressure- and heat-induced gels of food proteins. Agric. Biol. Chem. 54:183–189.
Oley, I., Verlinde, P., Hendrickx, M. E., and Van Loey, A. (2006). Temperature and pressure stability of
L-ascorbic acid and/or [6s] 5-methyltetrahydrofolic acid: A kinetic study. Eur. Food Res. Technol.
223:71–77.
Patazca, E., Koutchma, T., and Ramaswamy, H. S. (2006). Inactivation kinetics of Geobacillus stearo-
thermophilus spores in water using high-pressure processing at elevated temperatures. J. Food Sci.
71:M110–M116.
Penna, A., Rao-Gurram, S., and Barbosa-Canovas, G. V. (2007). Effect of the milk treatment on acidifica-
tion, physicochemical characteristics and probiotic cell counts in low fat yogurt. Milch-wissenschaft
62(1):48–51.
Reddy, N. R., Marshall, K. M., Morrissey, T. R., Loeza, V., Patazca, E. et al. (2013). Combined high pressure
and thermal processing on inactivation of type A and proteolytic type B spores of Clostridium botulinum.
J. Food Prot. 76(8):1384–1392.
Reddy, N. R., Tetzloff, R. C., Solomon, H. M., and Larkin, J. W. (2006). Inactivation of Clostridium botulinum
nonproteolytic type B spores by high pressure processing at moderate to elevated high temperatures.
Innov. Food Sci. Emerg. Technol. 3:169–175.
Saldo, J. (2016). Microbiology under pressure: How microorganisms are affected by pressure and how they
may cope with it? In An Introduction to High-Pressure Science and Technology (Ed.) J. M. Recio, J. M.
Menendez and A. O. D. L. Roza, pp. 335–352. Taylor & Francis Group, Boca Raton, FL.
Sedighi, M., and Jabbari, A. (2013). A new analytical approach for wire-wound frames used to carry the loads
of pressure vessel closures. J. Press. Vessel Technol. 135(6):061206-061207.
Skinner, G. E., Marshall, K. M., Morrissey, T. R., Loeza, V., Patazca, E. et al. (2014). Combined high pressure
and thermal processing on inactivation of type E and nonproteolytic type B and F spores of Clostridium
botulinum.J. Food Prot. 77(12):2054–2061.
Son, D. S. Hong, J. H., and Chang, S. H. (2012). Determination of the autofrettage pressure and estimation of
material failures of a Type III hydrogen pressure vessel by using finite element analysis. Int. J. Hydrogen
Energy 37:12771–12781.
Spain, I. L., and Paauwe, I. (1977). High Pressure Technology, Vol I. Marcel Dekker, New York.
Suzuki, A., Watanabe, M., Iwamura, K., Ikeuchi, Y., and Saito, M. (1990). Effect of high pressure treatment
on the ultrastructure and myofibrillar protein of beef skeletal muscle. Agric. Biol. Chem. 54:3085–3091.
Verbeyst, L., Oey, I., Van der Plancken, I., Hendrickx, M., and Van Loey, A. (2010). Kinetic study on the
thermal and pressure degradation of anthocyanins in strawberries. Food Chem. 123:269–274.
Vetter, G., Luft, G., and Maier, S. (2001). Design and construction of high-pressure equipment for research and
production. High Pressure Process Technology: Fundamentals and Applications (Ed.) A. Bertucco and
G. Vetter, pp. 141–242. Elsevier Science, Amsterdam, the Netherlands.
Weber, G. (1992). Protein Interactions, pp. 235–270. Chapman & Hall, New York.
Yaldagard, M., Mortazavi, S. A., and Tabatabaie, F. (2008). The principles of ultra high pressure technology
and its application in food processing/preservation: A review of microbiological and quality aspects.
Afr. J. Biotechnol. 7(16):2739–2767.
Yamazaki, A. (2006). Development of food products using high-pressure induced transformation (Hi-Pit).
Rev. High Press. Sci. Technol. 16:4–10.
Yordanov, D. G., and Angelova, G. V. (2010). High pressure processing for food preserving. Biotechnol
Biotechnol Equip. 24(3):1940–1945. doi:10.2478/v10133-010-0057-8.
ChaptEr 2
high hydrostatic pressure processing

of Cereals and pulses
Sajad Ahmad Wani and Pradyuman Kumar
CONtENtS
2.1 Introduction ............................................................................................................................ 11

2.2 Application of HHP on Cereals and Legumes ....................................................................... 12
2.2.1 High-Pressure Processing of Cereals ......................................................................... 13
2.2.2 Effect of HHP on Starch ............................................................................................. 14
2.2.3 Effect of HHP on Gluten ............................................................................................ 15
2.2.4 High-Pressure Processing of Legumes ....................................................................... 16
2.2.5 HHP Effect on Legume Proteins ................................................................................ 17
2.3 Denaturation of Proteins ......................................................................................................... 18
2.3.1 Effect of HHP on the Antinutritionals Present in Legumes ....................................... 19
2.3.2 Influence of HHP on the Constituents of Grains ........................................................20
2.3.3 Enzymes...................................................................................................................... 21
2.3.4 Microbial Inactivation ................................................................................................ 21
2.4 Future Aspects ........................................................................................................................ 23
References ........................................................................................................................................ 23
2.1 INtrODUCtION
The use of pressure as a process factor in the food industries is not a new concept. It was Hite
(1899) who witnessed that use of pressure could increase the shelf life of milk and other food prod-
ucts. However, it took more than a century before extensive research continued in this field. In order
to ensure prolonged shelf-life and food safety, previously high temperature was used as a method
of food processing. But, at the same time high temperature is generally known to cause detrimental
changes in processed foods in terms of nutritional as well as sensory attributes. Degradation of
various vitamins occurs due to high temperature, which also causes changes in colour as well as
flavour compounds. Changes in texture can also be noticed, often softening of food tissues, occur-
ring by high temperatures and as a result the food scientists go for chemical compounds to regain
firmness. These entire changes result in food products that are far from original fresh products.
This idea sets a new task for the food scientist, as new processing techniques need to be developed
and adopted in order to satisfy consumer demand. For that, novel technologies are presently under
extensive investigation. Among these, non-thermal processes are high hydrostatic pressure (HHP),
11
pulsed electric fields, high-intensity light pulses, and irradiation (San Martín et al., 2002). HHP
processing is a comparatively newer technique that has attracted attention of food scientists in the
past few decades. The main advantage of HHP processing is instant and homogenous transmittance
of pressure in the food product, leading to inactivation of microorganisms and shelf-life extension
of food products (Koutchma, 2014).
The use of HHP for food processing is growing within the food industry. Presently, one of
the main applications of HHP processes within the food industry is the extension of shelf-life
of foods, even though as the research advances other uses are foreseen. Some of these are sol-
ute diffusion processes (Lambert et al., 1999) (salting, sugaring), HHP-assisted freezing-thawing
processes (Chevalier et al., 1999), and changes in the functional properties of proteins and other
macromolecules (Stolt et al., 1999).
In HHP processing, food is submitted to high hydrostatic pressure (mostly between 100 and
1000 MPa) with the aim of destroying both spoilage microorganisms and pathogens and also at the
same time inactivating enzymes that may cause detrimental alterations (Farr, 1990; Hoover et al.,
1989). The application of this technology is being used for various types of foods and raw material
processing for obtaining outputs with innovative sensory and functional properties (Norton and
Sun, 2007; Welti-Chanes et al., 2005). Other advantages of HHP technology is that as it does not
use heat; as a result, the parameters such as sensory and nutritional properties of the food product
remain virtually unaffected, therefore yielding products with better quality as compared to those
processed with other methods like high heat (San Martín et al., 2002). Furthermore, HHP combined
with other treatments (e.g., low temperature) may increase its effectiveness and decrease the detri-
mental effects caused by HHP on foodstuff and other related ingredients. Such detrimental effects
include lipid oxidation in fish and rancid flavours in tomato juice (Pitts, 1999).
HHP processing has found an increasing acceptance for producing good quality foods in the
food industry. In 2013, a report by global market on the HHP-processed products market indicated
an achievement of about 3 billion dollars. Products based on fruits and vegetables are the major
product market for HHP processing, followed by products of meat, seafood, and fish. Globally the
HHP products and equipment market is dominated by North America, with the United States as the
largest market. The USA, as a leading adaptor of HHP technology, is followed by some European
countries (Spain, Germany, France, the Netherlands, Great Britain, and Italy), Asia (Japan, China,
and Taiwan), Australia, and New Zealand. The estimated HHP equipment market in 2013 was
350 million dollars and is likely to grow by 11% compound annual growth rate for the next five years
(Koutchma, 2014).
With the growth of HHP technology, it is necessary to understand the fundamental principles in
order to provide practical recommendation on how HHP produce can be developed and how HHP
can be used for particular operations, considering available scientific information. This involves the
phenomena of compression heating, effect of pressure on biological organisms and food constitu-
ents, microbial load of foodstuff, product properties, influence on quality factors, and suitability of
packaging materials. Simultaneously, the effects of critical processing factors that define the behav-
iour of pressure in combination with temperature need to be understood well from the efficacy and
productivity point of view, depending on the application.
2.2 appLICatION OF hhp ON CErEaLS aND LEGUMES
Globally, one of the main sources of calories in the human diet is cereals and legumes. They
provide important nutrients such as proteins and vitamins and are the important source of energy.
The grains of these cereals and legumes are generally thermally processed, which increases their
digestibility and eliminates allergens. Many of the studies conducted with HHP are related to
the processing of fruits, vegetables and meat. Only a few studies are available where HHP has
HiGH HyDrostAtiC Pressure ProCessinG oF CereALs AnD PuLses 13
been applied as a source of a non-thermal process to cereals and legumes for inactivation of
anti-nutritionals, while at the same time preserving its quality and food components. The purposes
of this article are to review the application of HHP processing on cereals and legumes and to
address its effects on various components and properties of those foods.
2.2.1 high-pressure processing of Cereals
Cereal grains are the essential sources of energy for humans (Cummings and Stephen, 2007).
They are important sources of carbohydrates, vitamins, and minerals. Other than the provision of
energy, the carbohydrates of cereal grains also have other effects on various physiological processes
vital for maintaining health and preventing diseases (Cummings and Stephen, 2007). They can
be used in combination with legumes to make a complete protein food. Cereals are consumed by
people globally. There are some people who have allergy to cereal grains. Such sensitivity is mani-
fested by intestinal disorders, weight loss, and anaemia that is due to intake of oats or wheat and
may involve the body’s interaction with the gluten. In order to avoid the grains that possess gluten,
a combination of individual proteins classified as glutenins and prolamins, and substituted by
others cereals grains like rice, corn, and millet can reduce such symptoms. Another disease related
to intolerance of cereal grains in humans is celiac disease, which is a malabsorption that may in
part be produced by an inability to handle grains possessing gluten. Furthermore, some nutritional
deficiencies, aspects related to diet, like protein-fat ratios, psychological factors, may contribute to
these intestinal symptoms of celiac disease as well.
There are a number of foodstuffs to which HHP has been successfully applied, such as oysters,
avocado puree, fruit juices, sliced jam, etc. These foods are present in the marketplace of various
countries (Norton and Sun, 2007; San Martín-Gonzalez et al., 2006). However, only limited lit-
erature is available about the usage of HHP in the discipline of cereals and cereal-based products.
Japanese sake and rice sake treated with HHP can be found in the Japanese market (Cheftel, 1995).
Allergenic proteins such as 7S globulins from rice grains are solubilized by HHP treatment. HHP
treatment 100–400 MPa releases 0.2–0.5 mg per gram of proteins in rice, and maximum amounts
were obtained in the pressure range 300–400 MPa (Figure 2.1). However, no change was observed
in other parameters such as shape, size, or colour of treated grains at moderate pressure. Reduction
Figure 2.1 effect of HHP on the release of proteins from rice grains: (a) 0–400 MPa for 30 min, (b) 300 MPa
for 0–120 min. (Adapted from Kato, t. et al., J. Agric. Food Chem., 48, 3124–3126, 2000.)
in microbial count and increasing protein digestibility has been found by HHP. Non-significant
changes in various nutritional constituents of grains such as vitamin A were observed, but vitamins
such as B1, B6, and C were well retained (about 85%). HHP has been applied for various other cere-
als such as wheat and barley flours. Effect of HHP on the activity of amylases and analysing new
textured products from doughs is being studied as well (Estrada-Girona et al., 2005).
Oat batters were treated for 10 min at 200, 300, 350, 400, or 500 MPa. Scanning electron
microscopy revealed that proteins and starch granules of oat batters were affected by HHP treat-
ment, changes becoming more evident at high pressures. As revealed from bright field microscopy,
HHP treatment of oat batter also affected proteins. A significant improved batter viscosity and
elasticity at high pressures was observed. Oat proteins, which are soluble in water, salt, and urea,
were affected by HHP treatment. Extent of protein modification was observed to be dependent on
the applied pressure treatment, and it was noticed that the results of HHP can be used to improve
the functionality of oat batters (Huttner et al., 2009).
Influence of HHP on wheat dough was examined by determining mechanical-, colour-, and
texture-related dough factors (adhesiveness, cohesiveness, stickiness, and hardness). An increase
in dough hardness and adhesiveness was observed by HHP treatment, but treatment time decreased
its stickiness. The microstructure of dough by scanning electron micrographs suggested that pres-
sure treatment higher than 50 MPa affected the proteins, but starch modification required higher
pressure levels. Treatment of HHP to yeasted doughs led to wheat breads with different appear-
ance and technological properties (brownish colour acquired by crumb and heterogeneous cell
gas distribution with increased hardness due to new crumb structure) (Barcenas et al., 2010).
There are a lot of studies describing the effect of HHP on particular cereal components properties
or model systems, such as gluten and starch (Apichartsrangkoon et al., 1998; Gomes et al., 1998;
Kieffer et al., 2007).
2.2.2 Effect of hhp on Starch
Starch is an essential biopolymers extensively used in the food industry. Moreover, being an
integral part of cereal grains, starch has been used as bulking agent, thickener, colloid stabilizer,
gelling agent, etc. Mostly modified starches are being used in the food industries and there is
a limited use of unmodified native starches in the food industry as the later lacks the stability
under shear, temperature, pH, and cool conditions (Liu et al., 2009). Common methods applied
for modification of starches for use in processed foods are physical and chemical methods caus-
ing changes in the physical and chemical properties of the native starch, thus improving its
functional properties (Regina et al., 1998). Among different methods of physical modification,
HHP processing is a non-thermal processing technology. It is an appropriate technique for the
production of minimally processed foodstuffs, and it has potential to be used for the develop-
ment of novel products (Farr, 1990).
As starch is the major constituent of cereals, it is the constituent of cereals most affected by HHP
processing. Use of HHP can be comparatively synergistic as an additive starch enhancement tech-
nique. HHP has been found to induce starch gelatinization and modification in about 25 starches.
The responses of different starches were different at high pressures depending on the source of
starch, range of pressure, pressurization temperature and time, solvents used, and concentration
of starch (Pei-Ling et al., 2010). In a study, starch samples were subjected to preselected HHP
of 200–1500 MPa and temperatures, and HHP was found to decrease gelatinization temperature.
Decrease in gelatinization temperature was found to be a nonlinear function of pressure showing
maximum effect at high pressures (Muhr and Blanshard, 1982). Starch gelatinization was observed
by HHP as revealed by differential scanning calorimetry, which started at 300 MPa and was almost
complete after treatment at 500 MPa. Extent of starch gelatinization was found to be dependent on
the applied pressure treatment, and it was found that the results at HHP can be used to improve the
functionality of oat batters (Huttner et al., 2009).
HHP processing affects the amorphous and ordered structure of starch and as a result causes
starch gelatinization as a function of applied pressure, temperature, time, amount, and kind of starch
(Pei-Ling et al., 2010). Starch swelling by HHP treatment takes place, keeping granule integrity.
HHP processing treatment affects the starches by modifying their microstructure and rheological
characteristics by different means as compared to thermally processed ones (Gomes et al., 1998;
Stolt et al., 2001). Extent of starch swelling depends on the kind of starch, level of pressure, and
treatment time (Stolt et al., 2001; Stute et al., 1996). Furthermore, the thermal properties of pres-
sure-treated starches indicate reduction in temperature of gelatinization and enthalpy; moreover, the
granular structure of the starches loses its crystallinity and as a result they become susceptible to
aggregation (Wang et al., 2008). X-ray examination revealed that HHP treatment converted starch
that showed the C-type X-ray pattern to the B-type-like pattern. HHP processing changed the shape
of starch granules and their surface appearance as per the SEM study. HHP processing at 600 MPa
for 30 min caused the entire gelatinization of mung bean starch (Li et al., 2011).
2.2.3 Effect of hhp on Gluten
Gluten is the protein constituent of wheat. Most of the product development depends on the
quality and quantity of gluten. Its presence is necessary in products such as various kinds of breads,
biscuits, and cakes. However, gluten has also been found to be responsible for certain kinds of
diseases such as celiac disease. Therefore, knowledge on the effect of HHP processing on gluten is
important.
The unique rheological characteristics of wheat dough are mainly due to gluten protein. Few
studies have revealed the effect of HHP on gluten properties. Lullien-Pellerin et al. (2001) have
studied the effect of HHP on protein conformation by using γ46 Gliadin. A change in conformation,
which causes reduction in the polarity of the environment of aromatic amino acids combined with
an increase in the hydrophobicity of the gliadin surface, was observed when HHP above 400 MPa
was applied. The changes detected were found to be reversible. Apichartsrangkoon et al. (1998,
1999) treated wheat gluten with HHP of 200–800 MPa and at a temperature of 20°C–60°C for 25
or 50 min and examined the products for structural modification by using stress rheometry, texture
profile analyser, and solubility in sodium dodecylsulphate solutions. Rheological properties were
found to be significantly affected by HHP and temperature. A change in gluten solubility by HHP
was observed even at low temperatures. However, disulphide cross-linking only became significant
once the samples were kept at 800 MPa for 50 min.
Application of low pressure (200 MPa) and temperature (30°C) to gluten has increased the ratio
of the ethanol-soluble fraction and reduced gluten strength. By increasing pressure and temperature,
a strong reduction in the ethanol-soluble fraction and thiol content of gluten was observed. In glia-
din types, cysteine containing α- and γ-gliadins, but not cysteine-free ω-gliadins, were sensitive to
pressure and were transferred to the ethanol-insoluble fraction. It was observed in the disulphide
peptides isolated from treated gluten that rearrangement and cleavage of disulphide bonds were
involved in pressure-induced reactions. HHP and temperature induced a significant strengthening
of gluten protein, and under HHP of 800 MPa and 60°C, losses in gluten cohesivity was observed.
Besides, the characteristics of isolated glutenin having relatively high thiol content were strongly
affected by HHP and temperature, compared to the effects on total gluten (Kieffer et al., 2007).
Glutenin after removal of gliadin from gluten was given HHP treatment at different temper-
atures for 10 min. The solubility of treated glutenin in sodium dodecylsulphate buffer strongly
reduced as HHP and temperature increased. Higher effects were noticed at 800 MPa and 70°C.
It was observed that the ratio of the sodium dodecylsulphate-soluble portion decreased to 1.9%
Figure 2.2 effect of pressure at various temperature conditions on proportion of sDs-soluble glutenin in
gluten. (Adapted from Kieffer, r. et al., J. Cereal Sci., 45, 285–292, 2007.)
(Figure 2.2) (Kieffer et al., 2007). From the overall observation and studies, it can be suggested that
HHP processing affects the cereals by affecting their main constituents such as starch and gluten
and, therefore, could be an alternative technique for obtaining novel textured cereal-based products
(Barcenas et al., 2010).
2.2.4 high-pressure processing of Legumes
Legumes are an important source of proteins, vitamins (particularly B-Vitamins), and minerals
such as calcium, magnesium, potassium, and zinc (Saha et al., 2009). The legumes are rich in lysine
but poor in methionine (Estrada-Giron et al., 2005) and cysteine (Han et al., 2007); therefore; they
can be mixed with cereals to improve the amino acid profile (Estrada-Giron et al., 2005) of the final
product. They occupy an essential place in human nutrition, mainly in those countries where the
consumption of meat is limited (Boye et al., 2010). Protein content of the legume is much higher
than the cereal crops and it is a cheap and easily available source of protein with low starch content
and highly resistant starch content. Legumes can be incorporated as a source of protein for the devel-
opment of various products such as bread, extruded product, and other products. Numbers of anti-
nutritional compounds have been found in legumes such as trypsin inhibitor, phytic acid, tannin, etc.
Use of HHP processing to legume has been found to be beneficial because of the destructive
effect of HHP treatment on the anti-nutritionals and modification of other constituents present in
legumes such as in-vitro protein digestibility and improvement in the texture of the final product.
A study on legume batters showed that HHP processing provokes variation on the rheology of
hydrated samples, mainly in softer batters, causing an increased solidity/stiffness. While analyz-
ing legume proteins that were extracted in different buffers indicated that pressures of >200 MPa
induced the creation of disulphide bonds, urea-insoluble complexes, and/or other strong aggregates
of protein, the degree of protein modification was dependent on the pressure applied (Angioloni and
Collar, 2013). Starch-water suspension of mung bean was subjected to an HHP treatment at 120,
240, 360, 480, and 600 MPa for 30 min and a decrease in swelling power, light transmittance, and
solubility was observed with HHP treatment. Peak, trough, final, and setback viscosity and pasting
temperature increased significantly, whereas a reduction in breakdown viscosity was observed with
increase in HHP treatment. Decrease in gelatinization temperatures and gelatinization enthalpy
upon HHP treatment was also observed (Li et al., 2011).
2.2.5 hhp Effect on Legume proteins
The protein is the main important component of legumes. It is, therefore, necessary to know the
effect of HHP processing on the protein content. However, a limited literature is available on the use of
HHP and its effect on legume proteins. However, there are several other studies showing the effect of
HHP treatment on the protein content of meat. Therefore, it can be assumed that the effect of HHP on
legume protein can be similar to the effect that occurs in proteins of other sources during research anal-
ysis. The effect of HHP processing has been found to change the structural and functional properties
of proteins irreversibly (Pfister et al., 2001; Winter, 2003). The sensitivity of protein structure to HHP
is mainly due to weakening of hydrophobic and electrostatic bonds. In such cases, protein aggregates
(e.g., casein micelles) separate into their subunits even at relatively low pressure (150 MPa). Among
the various structures of protein, secondary and tertiary structures are mostly affected by HHP at 4700
and 4200 MPa, respectively (Pfister et al., 2001). The β-sheet structures are more stable than α-helices.
On the basis of the nature of protein and the intensity of HHP used, denaturation may be reversible or
irreversible. Formation of new non-covalent bonds may lead to the development of new tertiary struc-
tures and aggregates. Covalent bonds remain naive up to 1000 MPa by HHP, except disulphide bonds
that can be broken and rearranged by thiol/disulphide exchange reactions. Various elements affecting
protein changes at HHP are time, temperature of treatment, additives, solvent, pH value, and protein
concentration. Because of the alteration in protein properties such as viscosity, water binding capac-
ity, gel formation, elasticity, solubility, and enzymatic activity, interest in HHP treatment has become
boundless in the area of protein-rich foods (Pfister et al., 2001).
The in-vitro protein digestibility of various legumes such as chickpeas, lentils, peas, and
soybeans increases by heating them in water. However, in-vitro protein digestibility of legumes is
inconsistent among legumes or comparatively less by soaking treatment, irrespective of the application
of HHP. Moreover, increase in the in-vitro protein digestibility of legumes is possible by soaking or
soaking with HHP before heat treatment (Han et al., 2007). Penas et al. (2011) determined the chem-
ical score of proteins of raw soybean and HHP-treated soybean (Table 2.1). It was revealed that the
table 2.1 Effect of hhp on the total protein and amino acids
(g/100 g protein) Content of Soybean Seeds
parameters raw Soybean hhp treated Soybean

protein (% dw) 45.0 44.3
Essential amino acids

Leu 6.10 6.18
Lys 4.32 4.40
Val 4.19 4.26
phe 4.13 4.17
Ile 4.12 4.22
tyr 2.91 2.67
trp 2.26 2.29
thr 2.89 2.52
his 2.03 1.93
Met 1.32 1.36
Cys 1.24 1.08
total Eaa 35.5 35.1
Cs 45.0 42.8
EaaI 72.4 70.9
(Continued)
table 2.1 (Continued) Effect of hhp on the total protein and

amino acids (g/100 g protein) Content
of Soybean Seeds
parameters raw Soybean hhp treated Soybean

Non-essential amino
acids
Glu 15.9 15.7
asp 7.25 7.26
arg 5.44 5.60
pro 4.26 4.25
ala 4.05 3.97
Ser 3.87 3.94
Gly 3.50 3.07
Source: Penas, e. et al., Food Chem., 125, 423–429, 2011.
amino acid methionine (Met) and cysteine (Cys) were the limiting amino acids in raw soybean and
were responsible for a chemical score value of 45. Similarly, chemical score values can be obtained
from the essential amino acid (EAA) content for soybean seeds in Tachiyutaka cultivar. A decrease
in chemical score value by HHP treatment of soy seeds by about 5% was observed (Takahashi et al.,
2003). The application of HHP to raw soybean led to clear chemical score reductions of total EAAs,
a value of 35.5 for raw soybean, and 35.1 for HHP-treated soybean. As far as the essential amino
acid index (EAAI) is concerned, the raw soybean presented a value of 72.4, which reduced by 1.2%
in HHP treated soybeans (Table 2.1) (Penas et al., 2011).
2.3 DENatUratION OF prOtEINS
Under normal conditions, it is experimentally difficult to observe the cold denaturation phenom-
enon for ordinary proteins above the freezing temperature of aqueous solvent. One of the strange
properties of water under pressure allows the liquid to undergo significant super cooling at HP. The
effect of such super cooling at HP has been used to study the denaturation of proteins at sub-zero
temperatures without freezing. A few years ago, cold denaturation of proteins began to attract sci-
entific attention. According to Le Chatelier’s Principle, reduction in temperature of a system should
lead to reduction in enthalpy and entropy. Therefore, as the highest ordered state of proteins was
supposed to be their native state, decrease in temperature could not be expected to lead to drastic
conformational changes. But, it was found experimentally that globular proteins undergo a change
of protein structure and a disruption at very low temperatures (Privalov, 1990). It was Jonas (1997)
who worked on the structure of pressure-denatured and pressure-assisted cold-denatured (cold dena-
turation at constant HP) ribonuclease A and compared them with the heat-denatured state. It was
concluded that pressure and pressure-assisted cold denatured enzymes have as much secondary
structure as enzymes that are heat-denatured, and the fact was emphasized that more work with other
proteins is important to determine whether the observed effect can be generalized. HP can support
the cold denaturation of carboxypeptidase Y as demonstrated by Kunugi et al. (1999). A schematic
diagram of protein denaturation is shown in Figure 2.3. Protein denaturation states by temperature
and pressure combinations are delimited by an ellipsoidal curve. A phase diagram of water is repre-
sented in this figure too (Figure 2.3). It is clear that cold denaturation is not possible at atmospheric
pressure due to the formation of ice at low temperature, but increase in pressure to cool water to
subzero temperatures, as the ice is not formed and cold denaturation takes place.
Figure 2.3 schematic diagram of protein denaturation. (Adapted from san Martín, M.F. et al., Crit. Rev. Food
Sci. Nutr., 42, 627–645, 2002.)
2.3.1 Effect of hhp on the antinutritionals present in Legumes
Although legumes are rich sources of various dietary constituents, the presence of several
antinutritional factors such as trypsin inhibitors, phytic acids, polyphenols, saponins, oxalates, and
haemagglutinin in them limits their use in various product developments. These antinutritional fac-
tors present in legumes must be removed or reduced to utilize the legume’s whole nutritional poten-
tial. Several studies have been conducted for removing these antinutritional factors (Chau et al.,
1997; Kakati et al., 2010; Rehman and Shah, 2005; Wani and Kumar, 2016). In legumes, the protein
digestibility is less than in casein or other animal proteins because of intrinsic structural aspects of
proteins as well as antinutritional factors of legumes (Park et al., 2010). Protease inhibitors hinder
pancreatic serine proteases and reduce protein digestibility (Guillamón et al., 2008). Proteins react
with tannins and form reversible or irreversible complexes (Lampart-Szczapa et al., 2003) that can
reduce protein digestibility (Park et al., 2010). Tannins were previously considered as antinutrition-
als. However, tannins that belong to polyphenols possess antiseptic, antioxidant (Lampart-Szczapa
et al., 2003), and anticarcinogenic characters (Rocha-Guzmán et al., 2007). Phosphorus, present as
phytic acid, cannot be digested properly by human beings. Because of a high degree of phosphoryla-
tion (IP4 to IP6), inositol can produce non-soluble complexes with polyvalent cations such as cop-
per, calcium, zinc, or iron that reduce their bioavailability (Máñez et al., 2002). Several attempts
have been made by using various processing techniques to decrease or remove antinutritionals, so
as to improve the nutritional potential by raising their digestibility and bioavailability. Cooking,
commonly done by boiling, can change nutrient constituents, decrease phytic acid and tannins,
and inactivate trypsin inhibitor and lectin activity (Kakati et al., 2010; Linsberger-Martin et al.,
2013; Raj Bhandari and Kawabata, 2006). However, increasing time and temperature of cooking
can decrease the nutritive value and lysine availability of legumes (Mubarak, 2005). There are also
other various processing techniques available that have been found to improve digestibility by the
inactivation of the antinutritionals mentioned here. HHP processing has been investigated and has
been found to successfully reduce these antinutritionals and improve digestibility. However, only a
few studies are available on the effects of HHP treatment on antinutritionals.
Recent studies showed that the use of HHP for processing has increased significantly because
of its high efficiency, speedy processing time, and economic feasibility. HHP could inactivate
table 2.2 phytic acid and total phenolic acid Content, trypsin Inhibitor activity and IVpD (In Vitro
protein Digestibility) in Untreated and pressure-treated peas and Beans
pressure (Mpa), trypsin Inhibitor

time, and activity (tIU/mg, In Vitro protein phytic acid
peas temperature dwb) Digestibility (%) (g/100 g dwb)
p09 100, 30 min, 20°C 0.93 79.9 1.08
p07 100, 30 min, 60°C 0.39 79.6 0.87
p11 100, 60 min, 20°C 1.06 78.6 1.04
p05 100, 60 min, 60°C 0.19 79.2 0.75
pCpa 350, 45 min, 40°C 0.75 80.7 0.86
p04 600, 30 min, 20°C 0.96 82.3 0.84
p06 600, 30 min, 60°C n.d. 83.3 0.79
p08 600, 60 min, 20°C 0.79 82.1 0.81
p10 600, 60 min, 60°C 0.02 85.8 0.74
Untreated pea — 1.36 82.3 1.15
Beans
B09 100, 30 min, 20°C 12.02 68.3 1.12
B07 100, 30 min, 60°C 11.21 68.8 1.15
B11 100, 60 min, 20°C 13.11 68.5 1.13
B05 100, 60 min, 60°C 10.14 67.9 1.01
BCpa 350, 45 min, 40°C 12.86 68.0 1.11
B04 600, 30 min, 20°C 13.30 69.0 1.10
B06 600, 30 min, 60°C 5.38 68.8 1.03
B08 600, 60 min, 20°C 14.00 68.9 1.03
B10 600, 60 min, 60°C 2.92 75.1 1.08
Untreated bean — 18.05 69.1 1.13
Source: Linsberger-Martin, G. et al., LWT-Food Sci. Technol., 51, 331–336, 2013.

a Mean of three centre points (PCP: mean of P01, P02, and P03; bCP, mean of b01, b02, and b03).
trypsin inhibitors, remove unpleasant flavours, and increase digestibility, solubility, and metabo-
lism of foodstuffs (Lin et al., 2006; Khattab and Arntfield, 2009; Mubarak, 2005; Zhang et al.,
2010). However, the consumption rate of legumes is quite low in Western nations. In Europe about
2.5 kg pulses per capita in a single year were consumed in 2007, among them, 1.2 kg was peas and
0.7 kg beans (FAO, 2011). Possible reasons may be the presence of antinutritional factors (Saha
et al., 2009) as well as long preparation and cooking time (Bede, 2007). Also, flatulence-causing
oligosaccharides such as stachyose, raffinose, and verbascose often prevent consumers from con-
suming legumes. To overcome these barriers the production of convenience products for quick
and easy final preparation by consumers is important. Oligosaccharides were reduced by HHP by
up to 48% in beans and 68% in peas, but reduction was lesser than the samples that were cooked.
Reduction in phytic acid by HHP was up to 11% in beans and 36% in peas (Table 2.2). Reduction
in total phenolics by HHP in some pressurized beans and peas as compared to untreated beans and
peas was observed. A decrease in trypsin inhibitor activity up to 84% in beans and 100% in peas
by HHP was also observed (Table 2.2). Protein digestibility rises by up to 8.7% in beans treated
at 600 MPa at 60°C regardless of time and by 4.3% in peas when treated at 600 MPa at 60°C for
60 min (Linsberger-Martin et al., 2013).
2.3.2 Influence of hhp on the Constituents of Grains
One of the important things that are taken into consideration while speaking of any processing
is the retention of nutritional properties of the product. It has been hypothesized that HHP does
not affect covalent bonds, and that various components such as vitamins and flavour and colour
compounds will remain unaffected after HHP treatment. As we know, cereals and legume grains
are good sources of vitamins like B1, B2, B3, B5, B6, and tocopherol, which are mainly present in
the scutellum and aleurone or both. Moreover, these grains are also important sources of minerals
such as potassium, phosphorous, magnesium, calcium, copper, and iron. However, no literature is
available, particularly on the use of HHP and its effect on vitamins and minerals in grains and their
sub products. But, the effect of HHP on the vitamin solutions and micronutrients in fruit systems are
somewhat unaffected. Therefore, one can assume that the effect of HHP on grains may be similar to
its effect on fruits and vegetables during research analysis. It was Sancho et al. (1999) who found the
degradation of water-soluble vitamins such as thiamine, pyridoxine, and ascorbic acid during HHP
processing and moderate thermal treatment in a multivitamin model system and in various food
materials. All these vitamins were chosen due to their sensitivity to physical factors that restrict
their use in conventional thermal systems. Degradation of thiamine during food processing leads
to development of volatile sulphur compounds, like thiazoles or furanes, which are responsible for
the development of aroma and overall quality of food. Ultra-HHP treatments showed no significant
effects on the loss of B6 and B1. The unprocessed model system had 1.475 mg/mL of thiamine and
3.725 mg/mL of pyridoxal; however, after the treatment with HHP at 600 MPa and 20°C for 30 min,
the system showed 1.468 mg/mL of thiamine (99.57% retention) and 3.794 mg/mL of pyridoxal
(101.84% retention).
A significant degradation was observed when vitamin C was subjected to HHP process-
ing in a multivitamin system. The intensity of the chosen treatment had no significant effect
on the degradation of ascorbate in the same multivitamin system. At a pressure of 200 MPa
20°C for 30 min it had a concentration of 1.832 mg/mL (87.83% retention), and after treatment
at 600 MPa and 20°C for 30 min it had a concentration of 1.847 mg/mL (88.58% retention)
(Estrada-Girona, 2005).
2.3.3 Enzymes
Influence of HHP treatment on the enzymes varied and depended on the pressure, origin of the
enzyme, temperature, time of processing, and nature of the substrates (Palou et al., 1999). Like other
important constituents of cereals and legume, effect of HHP treatment has not been studied much.
Gomes et al. (1998) observed increased enzymatic activity when a pressure treatment of 400 and
600 MPa was subjected to 10% wheat or barley flour slurries for various amylases of malt barley.
However, pressure treatments of 700 to 800 MPa showed a reduced activity. Food scientists have
suggested that a pressure range of 400 to 600 MPa could gelatinize starch granules, and modifica-
tion of enzyme active site could favour hydrolysis.
2.3.4 Microbial Inactivation
The effects of HHP on microbial inactivation will depend on a number of factors like kind of
microorganism, temperature, magnitude, time of the HHP treatment, and composition of suspen-
sion media or food. By considering these factors, appropriate pressure treatment should be applied
to assure inactivation of spoilage and pathogenic and vegetative cells of microorganisms present in
foods.
A study on the influence of HHP on wheat dough was examined by determining microbial
count (total aerobic mesophilic bacteria, molds, and yeasts). HHP reduced the endogenous micro-
bial population of wheat dough from 104 CFU/g to 102 CFU/g. Effect of HHP on the total aerobic
mesophilic bacteria, mold, and yeast is shown in Figure 2.4 (Barcenas et al., 2010). Decrease in
microbial growth after 1 min exposure at HHP treatment and no further significant reduction was
observed by increasing the treatment time, with the exception of samples treated at HP of 50 MPa,
Symbols: : 50 MPa; : 100 MPa; : 150 MPa; : 200 MPa; ×: 250 MPa.
Figure 2.4 influence of HHP processing on (a) total aerobic mesophilic bacteria and (b) on molds and yeasts
of wheat dough. (Adapted from barcenas, M.e. et al., LWT-Food Sci. Technol., 43, 12–19, 2010.)
which required longer treatment time. Generally, high pressure has been related to high microbial
inactivation, but such relationship was not found with the treatment time (Palou et al., 1998).
A decrease in the microbial population of tofu from an initial count of 5.54 × 104 cfu/g to 0.31,
1.56, or 2.38 log units was observed. The effectiveness of HHP treatment to decrease microbial
count at 400 MPa largely depends on the time of exposure (Prestamo et al., 2000) (Figure 2.5).
Prestamo et al. (2000) also proposed a reduction in psychrotrophs by HHP treatment of tofu by 2 log
units from an initial population of 1 × 103 cfu/g. Decrease in mesophilic microbes was by 1 log unit
from an initial number of 1.6 × 103 cfu/g. In the case of mold and yeasts, reduction from an initial
population of 2.64 × 103 cfu/g to 1 × 102 cfu/g was observed. Some of the microorganisms that were
found in tofu before HHP treatment—like Salmonella, Pseudomonadaceae, and gram-negative bac-
teria—were not identified after HHP processing. Microorganisms such as Listeria monocytogenes
and Yersenia enterocolitica, which are more resistant to HHP, were not found before and after HHP
Figure 2.5 Viable aerobic mesophilic population in tofu after treatment at 400 MPa and 5°C for 5, 30, and
45 min. (Adapted from Prestamo, G. et al., J. Agric. Food Chem., 48, 2943–2947, 2000.)
processing. However, certain microorganisms such as Bacillus cereus and Hafnia halvei were found
to be active even after HHP treatment of tofu.
Another factor than temperature and degree and period of HHP processing is the medium com-
position where microorganisms are dispersed, which influences significantly the effectiveness of
HHP processing on the decline of microbial population. Various constituents of food like fructose,
sucrose, glucose, and salts of food affect the resistance of microorganisms for pressure (Oxen and
Knorr, 1993). Such type of effect is often observed because food constituents seem to protect micro-
organisms from the effects of HP. Thus, a non-nutritive solution can decrease the pressure tolerance
of microorganisms. As mentioned above the microorganisms such as Hafnia halvei and Bacillus
cereus that remained active after HHP processing could describe the pressure protective effect that
food constituents exert over the extent of microbial reduction (Prestamo et al., 2000).
2.4 FUtUrE aSpECtS
HHP processing of cereal grains and legumes offers many benefits over traditional processing
methods that involve heat treatment for increasing the shelf life of stored products by lowering
the microbial population of spoilage microorganisms, inactivating undesirable food enzymes and
compounds. Furthermore, HHP technology also offers benefits that may be used to develop new
textured foodstuffs. Nowadays a number of products that can be processed by HHP are available.
However, a lot of research needs to be developed and optimal parameters of pressure, temperature,
and time need to be determined so as to yield a foodstuff with unique characteristics and good
nutritional properties and sensory acceptability. At the same time, the high cost of HHP process-
ing distinguishes it from other technologies; therefore, besides product research and development,
some studies concerning the cost of processing should be conducted at the same time to ensure the
successful application of this technology to more food products. In addition, research regarding
inactivation kinetics of microbes and unimportant enzymes is also required.
rEFErENCES
Angioloni, A., and Collar, C. (2013). Impact of high hydrostatic pressure on protein aggregation and rheologi-
cal properties of legume batters. Food Bioproc. Technol. 6: 3576–3584.
Apichartsrangkoon, A., Bell, A.E., Ledward, D.A., and Schofield, J.D. (1999). Dynamic viscoelastic behavior
of high-pressure-treated wheat gluten. Cereal Chem. 76: 777–782.
Apichartsrangkoon, A., Ledward, D.A., Bell, A.E., and Brennan, J.G. (1998). Physiochemical properties of
high pressure treated wheat gluten. Food Chem. 63: 215–220.
Barcenas, M.E., Altamirano-Fortoul, R., and Rosell, C.M. (2010). Effect of high pressure processing on wheat
dough and bread characteristics. LWT-Food Sci. Technol. 43: 12–19.
Bede, E.N. (2007). Effect of quenching on cookability of some food legumes. Food Control. 18: 1161–1164.
Boye, J., Zare, F., and Pletch, A. (2010). Pulse proteins: Processing, characterization, functional properties and
applications in food and feed. Food Res. Int. 43: 414–431.
Chau, C.F., Cheung, P.C.-K., and Wong, Y.S. (1997). Effects of cooking on content of amino acids and antinu-
trients in three Chinese indigenous legume seeds. J. Sci. Food Agric. 75: 447–452.
Cheftel, C.J. (1995). Review: High-pressure, microbial inactivation and food preservation. Food Sci. Technol.
Int. 1: 75–90.
Chevalier, D., Le Bail, A., and Chourot, J.M. (1999). High pressure thawing of fish (Whiting): Influence of the
process parameters on drip losses. LWT-Food Sci. Technol. 32: 25–31.
Cummings, J.H., and Stephen, A.M. (2007). Carbohydrate terminology and classification. Eur. J. Clin. Nutr.
61: S5–S18.
Estrada-Giron, Y., Swanson, B.G., and Barbosa-Canovas, G.V. (2005). Advances in the use of high hydrostatic
pressure for processing cereal grains and legumes. Trend. Food Sci. Technol. 16: 194–203.
FAO (2011). FAO statistics, food balance sheets. Accessed October 24, 2011. www.fao.org/faostat/en/#date/
FBS.
Farr, D. (1990). High pressure technology in the food industry. Trend. Food Sci. Technol. 1: 14–17.
Gomes, M.R.A., Clark, R., and Ledward, D.A. (1998). Effects of high pressure on amylases and starch in
wheat and barley flours. Food Chem. 63(3): 363–372.
Guillamón, E., Pedrosa, M.M., Burbano, C., Cuadrado, C., De Cortes Sánchez, M., and Muzquiz, M. (2008).
The trypsin inhibitor present in seed of different grain legume species and cultivar. Food Chem. 107:
68–74.
Han, I.H., Swanson, B.G., and Baik, B.K. (2007). Protein digestibility of selected legumes treated with ultra-
sound and high hydrostatic pressure during soaking. Cereal Chem. 84(5): 518–521.
Hite, B.H. (1899). The effect of pressure in the preservation of milk. West Virginia Agric. Exp. Bull. 58: 15–35.
Hoover, D.G., Metrick, C., Papineau, A.M., Farkas, D.F., and Knorr, D. (1989). Biological effects of high
hydrostatic pressure on food microorganisms. Food Technol. 43(3): 99–107.
Huttner, E.K., Bello, F.D. Poutanen, K., and Arendt, E.K. (2009). Fundamental evaluation of the impact of
high hydrostatic pressure on oat batters. J. Cereal Sci. 49: 363–370.
Jonas, J. (1997). Cold denaturation of proteins. ACS Symp. Ser. 676: 310–323.
Kakati, P., Deka, S., Kotoki, D., and Saikia, S. (2010). Effect of traditional methods of processing on the nutri-
ent contents and some antinutritional factors in newly developed cultivars of green gram [Vigna radiata
(L.) Wilezek] and black gram [Vigna mungo (L.) Hepper] of Assam, India. Int. Food Res. J. 17: 377–384.
Kato, T., Katayama, E., Matsubara, S., Omi, Y., and Matsuda, T. (2000). Release of allergic proteins from rice
grains induced by high hydrostatic pressure. J. Agric. Food Chem. 48: 3124–3126.
Khattab, R., and Arntfield, S. (2009). Nutritional quality of legume seeds as affected by some physical treat-
ments 2. Antinutritional factors. LWT-Food Sci. Technol. 42(6): 1113–1118.
Kieffer, R., Schurer, F., Kohler, P., and Wieser, H. (2007). Effect of hydrostatic pressure and temperature
on the chemical and functional properties of wheat gluten: Studies on gluten, gliadin and glutenin.
J. Cereal Sci. 45: 285–292.
Koutchma, T. (2014). Adapting High Hydrostatic Pressure (HHP) for Food Processing Operations. Elsevier,
London, UK.
Kunugi, S., Yamamoto, H., Makino, M., Tada, T., and Uehara-Kunugi, Y. (1999). Pressure-assisted cold-
denaturation of carboxypeptidase Y. Bulletin Chemic. Soc. Japan. 72: 2803–2806.
Lambert, Y., Demazeau, G., Largeteau, A., and Bouvier, J.M. (1999). Changes in aromatic volatile composi-
tion of strawberry after high pressure treatment. Food Chem. 67: 7–16.
Lampart-Szczapa, E., Siger, A., Trojanowska, K., Nogala-Kalucka, M., Malecka, M., and Pacholek, B. (2003).
Chemical composition and antibacterial activities of lupin seeds extracts. Nahrung/Food. 47: 286–290.
Li, W., Zhang, F., Liu, P., Bai, Y., Gao, L., and Shen, Q. (2011). Effect of high hydrostatic pressure on physico-
chemical, thermal and morphological properties of mung bean (Vigna radiata L.) starch. J. Food Eng.
103: 388–393.
Lin, J.T., Liu, S.C., Chen, S.L., Chen, H.Y., and Yang, D.J. (2006). Effects of domestic processing on steroidal
saponins in Taiwanese yam cultivar (Dioscorea pseudo japonica Yamamoto). J. Agric. Food Chem.
54(26): 9948–9954.
Linsberger-Martin, G., Weiglhofer, K., Thi Phuong, T.P., and Berghofer, E. (2013). High hydrostatic pressure
influences antinutritional factors and in vitro protein digestibility of split peas and whole white beans.
LWT-Food Sci. Technol. 51(1): 331–336.
Liu, H. S., Yu, L., Dean, K., Simon, G., Petinakis, E., and Chen, L. (2009). Starch gelatinization under pres-
sure studied by high pressure DSC. Carbohyd Polym. 75(3): 395–400.
Lullien-Pellerin, V., Popineau, Y., Meersman, F., Morel, M.-H., Heremans, K., Lange, R., and Balny, C.,
(2001). Reversible changes of the wheat g46 gliadin conformation submitted to high pressure and tem-
perature. Eur. J. Biochem. 268: 5705–5712.
Máñez, G., Alegría, A., Farré, R., and Frígola, A. (2002). Effect of traditional, microwave and industrial cook-
ing on inositol phosphate content in beans, chickpeas and lentils. Int. J. Food Sci. Nutr. 53: 503–508.
Mubarak, A. (2005). Nutritional composition and antinutritional factors of mung bean seeds (Phaseolus
aureus) as affected by some home traditional processes. Food Chem. 89(4): 489–495.
Muhr, A.H., and Blanshard, J.M.V. (1982). Effect of hydrostatic pressure on starch gelatinisation. Carbohyd
Polym. 2: 61–74.
Norton, T., and Sun, D.W. (2007). Recent advances in the use of high pressure as an effective processing tech-
nique in the food industry. Food Bioproc. Technol. doi:10.1007/s11947-007-0007-0.
Oxen, P., and Knorr, D. (1993). Baroprotective effect of high solute concentration against inactivation of
Rhodotorula rubra. LWT-Food Sci. Technol. 26: 220–223.
Palou, E., Lopez-Malo, A., Barbosa-Canovas, G.V., Welti-Chanes, J., and Swanson, B.G. (1999).
Polyphenoloxidase activity and color of blanched and high hydrostatic pressure treated banana puree.
J. Food Sci. 64(1): 42–45.
Palou, E., Lopez-Malo, A., Barbosa-Canovas, G.V., Welti-Chanes, J., Davidson, P.M., and Swanson,
B.G. (1998). High hydrostatic pressure come-up time and yeast viability. J. Food Prot. 61(12):
1657–1660.
Park, S.J., Kim, T.W., and Baik, B.K. (2010). Relationship between proportion and composition of albumins,
and in vitro protein digestibility of raw and cooked pea seeds (Pisum sativum L.). J. Sci. Food Agric.
90: 1719–1725.
Pei-Ling, L., Xiao-Song, H., and Qun, S. (2010). Effect of high hydrostatic pressure on starches: A review.
Starch/Stärke 62: 615–628.
Penas, E., Gomez, R., Frias, J., Baeza, M.L., and Vidal-Valverde, C. (2011). High hydrostatic pressure effects
on immunoreactivity and nutritional quality of soybean products. Food Chem. 125: 423–429.
Pfister, M.K.H., Butz, P., Heinz, V., Dehne, L.I., Knorr, D., and Tauscher, B. (2001). Influence of High Pressure
Treatment on Chemical Alterations in Foods: A Literature Review. BgVV, Pressestellen, 89 p.
Pitts, K. (1999). Developments in high pressure food processing. Food Aus. 51(5): 197.
Prestamo, G., Lesmes, M., Otero, L., and Arroyo, G. (2000). Soybean vegetable protein (tofu) preserved with
high pressure. J. Agric. Food Chem. 48: 2943–2947.
Privalov, P.L. (1990). Cold denaturation of proteins. Crit. Rev. Biochem. Mol. Biol. 25(4): 281–305.
Raj Bhandari, M., and Kawabata, J. (2006). Cooking effects on oxalate, phytate, trypsin and α-amylase inhibi-
tors of wild yam tubers of Nepal. J. Food Comp. Anal. 19(6): 524–530.
Regina, M., Gomes, A., Clark, R., and Ledward, D.A. (1998). Effects of high pressure on amylases and starch
in wheat and barley flours. Food Chem. 63: 363–372.
Rehman, Z.U., and Shah, W.H. (2005). Thermal heat processing effects on antinutrients, protein and starch
digestibility of food legumes. Food Chem. 91: 327–331.
Rocha-Guzmán, N.E., González-Laredo, R.F., Ibarra-Pérez, F.J., Nava-Berúmen, C.A., and Gallegos-Infante,
J.A. (2007). Effect of pressure cooking on the antioxidant activity of extracts from three common bean
(Phaseolus vulgaris L.) cultivars. Food Chem. 100: 31–35.
Saha, S., Singh, G., Mahajan, V., and Gupta, H.S. (2009). Variability of nutritional and cooking quality in bean
(Phaseolus vulgaris L.) as a function of genotype. Plant Food Hum. Nutr. 64: 174–180.
Sancho, F., Lambert, Y., Demazeau, G., Largeteau, A., Bouvier, M., and Narbonne, J.F. (1999). Effect of ultra-
high hydrostatic pressure on hydrosoluble vitamins. J. Food Eng. 39(3): 247–253.
San Martín, M.F., Barbosa-Cánovas, G.V., and Swanson, B.G. (2002). Food processing by high hydrostatic
pressure. Crit. Rev. Food Sci. Nutr. 42: 627–645.
San Martín-Gonzalez, M.F., Welti-Chanes, J., and Barbosa-Canovas, G. (2006). Cheese manufacture assisted
by high pressure. Food Rev. Int. 22: 275–289.
Stolt, M., Oinonen, S., and Autio, K. (2001). Effect of high pressure on the physical properties of barley starch.
Innov. Food Sci. Emerg. Technol. 1: 167–175.
Stolt, M., Stoforos, N.G., Taoukis, P.S., and Autio, K. (1999). Evaluation and modelling of rheological proper-
ties of high pressure-treated waxy maize starch dispersion. J. Food Eng. 40: 293–298.
Stute, R., Klingler, R.W., and Boguslawski, S. (1996). Effects of high pressures treatment on starches. Starch/
Starke. 48: 399–408.
Takahashi, M., Uematsu, Y., Kashiwaba, K., Yagasaki, K., Hajika, M., Matsunaga, R., Komatsu, K., and
Ishimoto, M. (2003). Accumulation of high levels of free amino acids in soybean seeds through integra-
tion of mutations conferring seed protein deficiency. Planta. 217: 577–586.
Wang, B., Li, D., Wang, L.J., Chiu, Y.L., Chen, X.D., and Mao, Z.H. (2008). Effect of high pressure homogeni-
zation on the structure and thermal properties of maize starch. J. Food Eng. 87(2): 436–444.
Wani, S.A., and Kumar, P. (2016). Effect on nutritional, antioxidant and microstructural characteristics of
nutritionally enriched snacks by extrusion cooking. J. Food Proc. Preserv. 40(2): 166–173.
Welti-Chanes, J., Lopez-Malo, A., Palou, E., Bermudez, D., Guerrero-Beltran, J.A., and Barbosa-Canovas, G.V.
(2005). Fundamentals and applications of high pressure processing to foods. In: Novel Food Processing
Technologies, pp. 157–181. Barbosa-Canovas, G.V., Tapia, M.S., and Cano, M.P., Eds., Marcel Dekker/
CRC Press, New York.
Winter, R. (2003). Advances in High Pressure Bioscience and Biotechnology II. Springer, Berlin, Germany.
Zhang, M., Chen, H., Li, J., Pei, Y., and Liang, Y. (2010). Antioxidant properties of tartary buckwheat extracts
as affected by different thermal processing methods. LWT-Food Sci. Technol. 43(1): 181–185.
ChaptEr 3
Effect of high-pressure processing on

Selected Food processing Operations
Jincy M. George and Navin Kumar Rastogi
CONtENtS
3.1 Introduction ........................................................................................................................... 27

3.2 Effect of High Pressure on Infusion ..................................................................................... 29
3.3 Effect of High Pressure on Extraction .................................................................................. 30
3.3.1 Fruits, Vegetables, and Their By-products ............................................................... 30
3.3.2 Cereals ...................................................................................................................... 31
3.3.3 Herbs and Roots ........................................................................................................ 32
3.3.4 Seafood and By-products .......................................................................................... 33
3.3.5 Other Plant Products ................................................................................................. 33
3.4 Effect of High Pressure on Dehydration ............................................................................... 33
3.5 Effect of High Pressure on Rehydration ............................................................................... 35
3.6 Effect of High Pressure on Freezing ..................................................................................... 36
3.7 Effect of High Pressure on Thawing .................................................................................... 37
3.8 Effect of High Pressure on Frying ........................................................................................ 38
3.9 Benefits and Limitations of HPP .......................................................................................... 38
3.10 Concluding Remarks............................................................................................................. 39
References ........................................................................................................................................ 39
3.1 INtrODUCtION
Nutritional quality, microbiological safety, and pesticide residues in food commodities are
major consumer concerns (Miles et al., 1999). High-pressure processing (HPP) has emerged as a
healthier option for the consumers. These new dimensions of food processing give food processors
the opportunity to process foods with cleaner ingredients and fewer additives (Balasubramaniam
and Farkas, 2008). Preservation of foods by HPP is one of the novel and attractive alternatives as
it tends to have minimal effects on the food quality (Cullen et al., 2012). However, the preserva-
tive effect of HPP was discovered way back in the eighteenth century and it has been used success-
fully in the chemical, metallurgical, and plastic industries for decades, but it was only in the late
1980s that commercial benefits became available to the food processing industries. Hite (1899)
investigated the application of high pressure for preserving milk, and later it was used for the
preservation of fruits and vegetables (Hite et al., 1914). The first high-pressure-processed foods
27
were introduced to the Japanese market in 1990 by Meidi-ya Company, who have been marketing
a variety of jams, jellies, and sauces packaged and processed without application of heat (Thakur
and Nelson, 1998). Later, a US-based company called M/s Fresherized introduced high-pressure
preserved guacamole dip.
In addition to food preservation, high-pressure treatment can result in food products acquir-
ing novel structure, and therefore it can be used to develop new products as well as to increase the
functionality of certain ingredients (Hayashi, 1990). Based on the operating parameters, the cost
of HPP is typically lower (~US$ 0.05–0.5 per liter or kilogram) as compared to thermal processing
(Thakur and Nelson, 1998; Balasubramaniam, 2003). During high-pressure treatment, the food
product to be treated is placed in a pressure chamber capable of sustaining the desired pressure;
the sample to be pressurized is then submerged in a liquid, which acts as the pressure-transmitting
medium. The effect is similar to subjecting the food to a depth of 60 km in an ocean. Water or media
containing castor oil, silicone oil, ethanol, or glycol are used as the pressure transmitting medium
(Hogan et al., 2005). There are two general scientific principles based on the use of HPP in the food
processing industry. The first is Le Chatelier’s Principle, which states that when a system at equi-
librium is disturbed the system responds in a way that tends to minimize the disturbance (Pauling,
1964). Secondly, the isostatic rule states that pressure is instantaneously and uniformly distributed,
whether the sample is in contact directly with the pressurizing medium or sealed hermetically
(Olsson, 1995).
The early applications of high pressure were hampered due to unavailability of suitable equip-
ment. However, recent progress in design and development of equipment has ensured worldwide
recognition of the potential for such a technology in the food sector (Galazka and Ledward, 1995;
Gould, 1995; Balci and Wilbey, 1999). Today, high-pressure technology is recognised to have the
promise of developing a variety of products, simultaneously showing the potential for creating a
new generation of value-added food products. An increasing number of products produced using
this kind of non-thermal technology are now entering the food market. These include jams, fruit
dressings, yoghurt, fruit juices, dairy products, and non-frozen tropical fruits. In general, high-
pressure technology can supplement conventional thermal processing for reducing microbial load,
or substitute the use of chemical preservatives (Rastogi et al., 1994). A number of attempts have
been made to use HPP instead of high temperatures to inactivate food-spoiling microorganisms,
food-borne pathogens (Rendueles et al., 2011), and undesired food enzymes (Farr, 1990) while
maintaining all the quality and safety parameters of the products (Ferrari et al., 2010; Zhang et al.,
2012). The required pressure treatment for microbiologically safe and stable products is dependent
on the target microorganism to be inactivated. Bacterial vegetative cells, yeasts, and moulds are
sensitive to pressures between 200 and 700 MPa; bacterial spores may survive pressurization above
1000 MPa (Sale et al., 1970; Cheftel, 1992). HPP and additional hurdles like low pH were also used
synergistically in the case of fermented foods to prolong the shelf life (George and Rastogi, 2016).
The food processing sector uses various mass transfer operations for various processes. The
diffusion phenomenon governs these operations. A number of methods have also been tried to
improve mass transfer operations, including subjecting the food material to ultra-high hydrostatic
pressure (Rastogi and Niranjan, 1998) or high-intensity electrical field pulses (Amami et al., 2006)
prior to osmotic dehydration or applying ultrasound (Rastogi, 2011), partial vacuum (Rastogi and
Raghavarao, 1996; Fito et al., 2001), or centrifugal force (Azuara et al., 1996) during osmotic
treatment. HPP causes permeabilization of cell membrane or structural transformations, thereby
reducing the processing time for various operations (Farr, 1990; Dornenburg and Knorr, 1993;
Eshtiaghi et al., 1994; Rastogi et al., 1994). It is considered as a promising alternative to conven-
tional processing methods by the food manufacturers across the globe. Presently, there are a number
of high-pressure-processed products launched in the global food market by countries like Spain,
Japan, France, Italy, Portugal, U.K., U.S., and Canada (Hugas et al., 2002). The following section
outlines the effect of HPP on selected food processing operations.
eFFeCt oF HiGH-Pressure ProCessinG on seLeCteD FooD ProCessinG oPerAtions 29
3.2 EFFECt OF hIGh prESSUrE ON INFUSION
The demand for functional food is rising in emerging markets due to its health-promoting ben-
efits and not merely for its basic nutrition. These foods were targeted at preventing health deteriora-
tion and improving overall wellbeing. High-pressure pre-treatment of solid foods was explored as
a promising technique for infusion of bioactive compounds. There are numerous studies carried
out on the infusion of small molecules (salt, sugar) and large-sized molecules such as polyphe-
nols (>500 Da) into food matrices. Application of high pressure has been reported to accelerate
the diffusion of components into the food (Rastogi and Niranjan, 1998). The application of high
hydrostatic pressure to fruits and vegetables affects cell structure, making the cell more permeable,
which in turn enhances the uptake of biologically active substances from the surrounding solution
(Rastogi et al., 2007). George et al. (2016) have studied the infusion of anthocyanin from Garcinia
indica Choisy fruit in apple slices (Figure 3.1). It was revealed that high-pressure pre-treatment
could be a feasible technique for infusion of the bioactive compounds without altering the natural
matrix. Mahadevan et al. (2014) have shown an enhanced infusion of natural antioxidant (quercetin)
into frozen-thawed cranberries using high hydrostatic pressure processing. An amount that could
be infused in 10 min under high pressure needed at least 3 h under atmospheric condition (control),
while the amount of quercetin infused into high-pressure-processed cranberries was demonstrated
to be three times that of the control.
Similarly, Tola and Ramaswamy (2013) studied HPP acidification of low-acid foods like car-
rots. HPP acidification provided more uniform and rapid acidification compared to conventional
methods. Duvetter et al. (2005) and Fraeye et al. (2010) demonstrated that pressure-assisted infu-
sion of pectin methylesterase and calcium chloride in strawberries was capable of improving the
firmness of the strawberries. Rastogi et al. (2008, 2010) demonstrated that calcium infusion with
mild heating and application of high pressure could be used as a method for the improvement in
the texture of carrot during thermal- as well as pressure-assisted thermal processing. Sopanangkul
et al. (2002) indicated high-pressure treatment resulted in an eight-fold increase in the diffusion
coefficient of sucrose into potato cylinders as compared to ambient conditions. Similarly, Rastogi
and Niranjan (1998) observed two-fold increase in sucrose gain during osmotic dehydration of
high-pressure-treated pineapples.
1400
1200
Anthocyanin conc. (mg/100g)
1000
800
600
Control
400 50 MPa
150 MPa
200 250 MPa
350 MPa
0
0 1.0 2.0 3.0 4.0 5.0
Immersion time (h)
Figure 3.1 effect of high-pressure processing (50–350 MPa/10 min) on infusion of anthocyanin in apple slices.
(From George, J.M. et al., Innov. Food Sci. Emerg. Technol., 33, 100–107, 2016. With permission.)
3.3 EFFECt OF hIGh prESSUrE ON EXtraCtION
Besides being an attractive alternative processing tool to enhance the shelf life and reduce the
microbial load, high-pressure processing has the potential to alter the extractability of food com-
pounds (Jung, 2016). The use of high-pressure treatment in the extraction process creates novel
and interesting methodologies, which are often complementary to conventional extraction methods
(soxhlet extraction, soaking, maceration, water percolation), thereby enhancing the yield and reduc-
ing the processing time (Chemat et al., 2012, 2015; Rombaut et al., 2014). The use of HPP in extraction
of bioactive ingredients like flavanoids (Yan et al., 2008; Prasad et al., 2009a; Plaza et al., 2011),
polyphenols (Seo et al., 2011; Santos et al., 2013), saponins (Shouqin et al., 2007; Chen et al., 2009),
proteins (Altuner et al., 2012), natural colorants (Xi, 2006; Corrales et al., 2009; Strati et al., 2015;
Li et al., 2016), and polysaccharides (Guo et al., 2012) has been successfully carried out by various
researchers.
3.3.1 Fruits, Vegetables, and their By-products
The processing of foods generates an enormous quantity of by-products. Disposal of these by-
products adds cost to the food processor as well as creating a potential negative impact on the envi-
ronment. The recovery and use of by-products can lessen the waste disposal problems. The use of
high pressure for extraction of corilagin, lignin, and polysaccharides from longan fruit pericarp was
compared with other extraction methods wherein high-pressure-assisted extraction exhibited the
higher efficiency (Prasad et al., 2009b; Yang et al., 2009). Xi (2006) demonstrated that the higher
recovery (92%) of lycopene from tomato waste was obtained by performing high-pressure-assisted
extraction (500 MPa for 1 min).
Two of the most interesting agri-food wastes that contain phenolic species are grape marc and
olive pomace. Grape marc production (skins and seeds) is approximately 20–25 kg for every 100 kg
of grapevine produced (Passos et al., 2013), and olive pomace is made up of the solid residue of
olive pulp and seeds (Olea europaea L.), which is obtained during olive oil production. The extrac-
tion of high value-added compounds from these two agri-food wastes can play an important role
both in the economic sustainment of winemaking and olive oil production and in the reduction
in this industrial waste’s environmental impact. Paini et al. (2016) have also demonstrated that
high-pressure extraction of grape marc and olive pomace yielded higher phenolic extraction.
Corrales et al. (2009) found that high-pressure extraction (600 MPa, 70°C) of anthocyanin from
red grape skin resulted in higher antioxidant capacity as compared to control and the extraction yields
were also three-fold higher. Briones-Labarca et al. (2015) indicated that the antioxidant capacity of
Chilean papaya seeds increased to 129.3%, 242.7%, and 272.8% for high-pressure-treated samples
at 500 MPa for 5 min, 10 min, and 15 min, respectively, as compared to conventional extraction
(Figure 3.2). Casquete et al. (2014) indicated less intense pressure treatments should be applied
(300 MPa, 10 min) in the case of citrus peel to increase the phenolic content and antioxidant activ-
ity. Intense pressure treatments (500 MPa, 10 min) resulted in a decrease in the phenolic content and
antioxidant activity. Guo et al. (2012) indicated that the extraction yield and viscosity of pectin from
orange peel both were found to be higher for high-pressure extraction (500 MPa, 55°C for 10 min)
(Figure 3.3).
Perez-Lopez et al. (2016) indicated that the combination of HPP plus Ultraflo® L on Okara
improved the solubility of the dietary fiber, making it more suitable to be used for developing func-
tional foods. Strati et al. (2015) demonstrated that high-pressure-assisted extraction led to higher
extraction yields as compared to conventional solvent extraction process performed at ambient pres-
sure for 30 min. Plaza et al. (2012) reported that high-pressure treatment (200 MPa/25°C/6 min)
of astringent persimmon fruit at maturity stage showed the increase in carotenoid extractability
compared to the control samples.
140
120 gC
fB
DPPH (μmol TE g-1 seeds)

100
80 eA
60 d3
c2
40 a b1
20
0
CE UAE/ UAE UAE HHPE HHPE HHPE
5 min 10 min 15 min 5 min 10 min 15 min
Extraction method
Figure 3.2 Antioxidant capacity as DPPH (mmol te g−1 seed) from Chilean papaya seeds extracted by conven-
tional extraction (Ce), uAe (ultrasound) and HHPe (high hydrostatic pressure extraction). Different
letters above the bars indicate significant differences between mean values (p ≤ 0.05). (From
briones-Labarca, V. et al., Lebensm. Wiss. Technol., 60, 525–534, 2015. With permission.)
20 16
35°C 35°C
45°C 45°C
16 55°C 12 55°C
Yield of pectin (%)
Viscosity (mPas)
12 8
8 4
(A) (B)
4 0
300 350 400 450 500 300 350 400 450 500
Pressure (MPa) Pressure (MPa)
Figure 3.3 effect of pressure and temperature combination on the extraction yield and viscosity of pectin
extracted by high pressure from orange peel. HPP conditions: pressure from 300 to 500 MPa,
temperature from 35°C to 55°C, pressure-holding time of 10 min, liquid–solid ratio of 50:1. (From
Guo, X. et al., Carbohydr. Polym., 88, 441–448, 2012. With permission.)
3.3.2 Cereals
Kim et al. (2015) studied the effect of high-pressure treatment on the extraction of functional
components from germinated rough rice. Germination of rough rice was carried out at 37°C for
6 days and subjected to a high hydrostatic pressure treatment at 30 MPa for 24 h (HP24) and 48 h
(HP48). Germinated rough rice without high-pressure treatment (HP0), HP24, and HP48 were ana-
lyzed for their functional components. The γ-oryzanol content after HP24 and HP48 treatment
was 40.90 mg/100 g (after 6 days of germination) and 40.32 mg/100 g (after 1 day of germination),
respectively. These results revealed the efficacy of HPP to enhance the bioactive content in germi-
nated rough rice (Figure 3.4). Park et al. (2016) indicated that enzymatic hydrolysis by celluclast
60
HP0 HP24 HP48
γ-Oryzanol contents (mg/100g, dry basis)

50
Aa Aa
Ca Bca Ba Bb
Bb Ba Da Cb Ab
40 Bb BCb
Ea Eb Cc Dc Cc
Fb Dc
30
Ec
20
10
0
0 1 2 3 4 5 6
Germination period (Day)
Figure 3.4 Change in the γ-oryzanol content of germinated rough rice in high-pressure treatment (HPt). HP0;
without HPt, HP24 and HP48; applied HPt at 30 MPa during 24 h and 48 h, respectively. Values
are mean ± sD of 3 replicates. Different capital and small letters in the same items indicate a
significant difference (p < 0.05) among different germination periods and pressure treatment of
germinated rough rice, respectively. (From Kim, s. et al., Food Chem., 166, 86–92, 2015. With
permission.)
(0.5% w/w) in combination with HPP treatment (500 MPa) improved the extraction yield of tricin
(32.9 mg/kg rice hull), as compared to the traditional solvent extraction (14.7 mg/kg rice hull) or
HPP treatment alone (19.7 mg/kg rice hull). Higher levels of xanthohumol content in beer wort were
obtained by HPP treatments. Xanthohumol is a bioactive compound that has anti-carcinogenic,
antioxidant, anti-inflammatory, and anti-infective properties (Santos et al., 2013).
3.3.3 herbs and roots
Zhang et al. (2012) investigated the effect of high pressure on the extraction of bioactive com-
pounds such as baicalin and baicalein from S. baicalensis. The results revealed that, at the optimum
extraction conditions (Pressure 142.60 MPa, ethanol concentration 60.54%, solid-liquid ratio 1:35),
the extraction efficiency of baicalin and baicalein was 16.04 and 1.65%, respectively. Kim et al.
(2014) optimized the operating conditions for high-pressure extraction to maximize the extraction
yield of kirenol from Siegesbeckia orientalis L. (a Korean herb). The optimal processing conditions
(pressure 320 MPa, 5 min, ethanol concentration 18%, and feed-to-solvent ratio 1:76) resulted in the
highest extraction yield of kirenol (85.90%). High-pressure-assisted extraction of salidroside from
the herb Sedum (Rhodiola sachalinensis) at room temperature showed higher efficiency (0.40% in
3 min) as compared to that of ultrasonic extraction (0.29% in 30 min) and reflux extraction (0.30%
in 120 min) (Bi et al., 2009).
Shouqin et al. (2007) found that the yields of ginsenosides from Panax ginseng increased lin-
early within a pressure range of 100–500 MPa. When the pressure was higher than 500 MPa, the
yields increased only marginally (Figure 3.5). Chen et al. (2009) reported that high-pressure extrac-
tion of ginsenoside (200 MPa, 60°C, 5 min) gave a higher extraction yield as compared to soxhlet
extraction (90°C). Lee et al. (2011) demonstrated that high-pressure extraction of fresh and red
ginseng was found to be more effective than thermal extraction.
7.55
7.50
Yield of ginsenosides (%)

7.45
7.40
7.35
7.30
7.25
7.20
100 200 300 400 500 600
Pressure (MPa)
Figure 3.5 the yield of ginsenosides (%) as a function of pressure. (From shouqin, Z. et al., J. Food Eng., 79,
1–5, 2007. With permission.)
3.3.4 Seafood and By-products
Rodrigues et al. (2016) indicated that the extraction yield of Sargassum muticum (Yendo)
Fensholt (an edible seaweed) varied between 32% and 40.4% independently of the extraction condi-
tions (5–5.5 min, 300 MPa) or solid/liquid ratio, resulting in average increase of 3.6 to 4.8 times for
total sugars and sulfated sugars, as compared to conventional methods of extraction. Li et al. (2016)
indicated that the optimized conditions for pressurized extraction (200 MPa for 5 min; ethanol
solvent liquid to solid ratio of 20 mL/g) of astaxanthin from dried shrimp waste were higher and
extraction time less as compared to conventional solvent extraction.
3.3.5 Other plant products
Altuner et al. (2012) investigated the effect of high pressure on the extraction of protein from
Cedrus atlantica pollens. The total amount of proteins extracted by high pressure (220–330 MPa)
varied between 18.02 and 25.93 μg/mL, while it was 1.95 μg/mL in 24 h by the conventional extrac-
tion method. Altuner et al. (2014) also investigated the effect of high pressure on the extraction of
proteins from Betula pendula pollens. The results suggested that the amount of protein extracted
at 200 MPa for 5 min was similar to the conventional method of extraction that lasted for 24 h.
Hu et al. (2015) indicated that the extraction yield of chlorogenic acid from L. japonica flower buds
was higher for high pressure (400 MPa, 2 min, 30°C) extraction than for other methods.
3.4 EFFECt OF hIGh prESSUrE ON DEhYDratION
Fruits and vegetables pre-treated with high pressure enhance the drying rates, thereby reducing
the drying time (Ohlsson and Bengtsson, 2002). Different pretreatments such as hot water blanching
(Al-Khuseibi et al., 2005; Raikham et al., 2015), HPP (Yucel et al., 2010; Vega-Galvez et al., 2011),
pulsed electric field (Ade-Omowaye et al., 2003; Won et al., 2015), ultrasound (Mothibe et al., 2011;
Kowalski et al., 2015) and osmosis treatments (Pan et al., 2003) were used as pretreatments by many
researchers in order to reduce the drying time (Jangam, 2011). However, treatments associated with
high temperatures like hot water and steam blanching may affect the compounds associated with
sensory and nutritional quality attributes (Al-Khuseibi et al., 2005). Oliveira et al. (2015) investi-
gated the effect of HPP (600 MPa, 5 and 30 min) on drying characteristics of cocoyam, peruvian
carrot, and sweet potato cylinders. The results showed that HPP could be used as an interesting
tool for the treatment of tubers, leading to the increase in its drying rate and consequently reduc-
ing the processing time (Figure 3.6a–c). Hulle and Rao (2016) studied the effect of high-pressure
1.2
Dimensionless moisture content
1.0
0.8
0.6
0.4
0.2
0
0 200 400 600 800 1000 1200 1400 1600
Drying time (min)
1.2 Control 600 MPa/ 5 min 600 MPa/ 30 min

1.0
0.8
0.6
0.4
0.2
0
0 200 400 600 800 1000 1200 1400 1600
Drying time (min)
Control 600 MPa/ 5 min 600 MPa/ 30 min
1.2
1.0
0.8
0.6
0.4
0.2
0
0 200 400 600 800 1000 1200 1400 1600
Drying time (min)
Control 600 MPa/ 5 min 600 MPa/ 30 min
Figure 3.6 Drying curve of the control and HPP-processed sweet potato (a), cocoyam (b), and Peruvian carrot
(c) at 50°C. results are expressed as mean ± standard deviation. the a, b, c letters mean significant
difference (p ≤ 0.05) among the process for the same tuber. (From oliveira, M.M. et al., Innov. Food
Sci. Emerg. Technol., 31, 45–53, 2015. With permission.)
pre-treatment (300–500 MPa, 5–15 min) on dehydration characteristics of aloe vera cubes. The
results showed that high-pressure pre-treatment resulted in higher drying rates leading to a reduc-
tion in drying time as compared to control. The high-pressure treatment also enhanced the firmness
of aloe vera cubes. Janowicz and Lenart (2015) indicated higher density and shrinkage in convec-
tive dried apple cubes after high-pressure treatment. Yucel et al. (2010) indicated that high-pressure
pre-treatment (100–300 MPa) of beans also resulted in higher drying rates. Similarly, high-pressure
pre-treatment (400 MPa, 10 min) of red bell pepper (Capsicum annuun) resulted in higher drying
rates, which can be used as an alternative to chemical (NaOH and HCl) pre-treatments, thereby
minimizing environmental pollution (Ade-Omowaye et al., 2001). Eshtiaghi et al. (1994) have
studied the effect of different pre-treatments on the drying characteristics of green beans, carrots,
and potatoes. It was observed that pressure-treated samples had higher drying rates compared to
untreated samples.
Osmotic dehydration is an age-old technique that is commonly used for partial dehydration
and diffusion of bioactive compounds into the solid food matrix. The rate of mass transfer dur-
ing osmotic dehydration is generally low. Combined osmotic dehydration and high-pressure
pre-treatment is an innovative processing technique that can enhance the rate of water loss and
solid gain (Nunez-Mancilla et al., 2011). The application of high-pressure treatment also resulted
in an increased diffusion of moisture content (four-fold) during osmotic treatment of pineapples
(Rastogi and Niranjan, 1998). Verma et al. (2014) showed that high pressure as a pretreatment
for osmotic dehydration of banana slices resulted in enhanced drying rates. Nunez-Mancilla
et al. (2014) investigated the combined effect of osmotic dehydration and HPP in the range
from 100 to 500 MPa on quality parameters (firmness) of sliced strawberries stored at 5°C for
14 days. HPP resulted in a notable decrease in firmness compared to control sample as well as an
increase in the polysaccharide content with pressure. Dermesonlouoglou et al. (2017) indicated
that combined osmotic dehydration and high-pressure treatment of tomatoes extended their shelf
life to 181 days at 5°C. Leng et al. (2013) demonstrated that water uptake (or total weight) of
chicken breasts in 4% sodium chloride solution was found to increase with treatment pressure
up to 150 MPa; further increase in pressure up to 300 MPa resulted in a decrease in uptake of
water. Lakshmanan et al. (2007) indicated that fresh salmon had less water-holding capacity
than smoked salmon and a pressure of 150 MPa for 10 min caused a 2% higher water-holding
capacity of smoked salmon. Grossi et al. (2011) investigated the effect of high-pressure treat-
ment (500–600 MPa/1 s to 9 min/40°C–60°C) on pork sausages, which significantly increased
Young’s modulus and Hencky’s strain values. Perez-Won et al. (2016) studied the simultaneous
application of osmotic dehydration and high pressure for drying of red abalone (Haliotis rufe-
scens) slices. During the process, the drying time was shown to be reduced by high-pressure
pretreatment (350 and 550 MPa).
3.5 EFFECt OF hIGh prESSUrE ON rEhYDratION
Most of the ready-to-eat food products before consumption are rehydrated. The challenge asso-
ciated with the use of dehydrated foods is the loss of solids during rehydration. Rastogi et al.
(2000) studied the variation of moisture and solid content during rehydration of dried pineapples
subjected to high-pressure treatment before a two-stage drying process consisting of osmotic dehy-
dration and finish-drying at 70°C. As a result of cell permeabilization caused by the high pressure,
treatment led to the absorption of more water in comparison with control. At the same time, loss
of solute during rehydration was also reduced, probably due to structural changes induced during
high-pressure pre-treatment. Ramaswamy et al. (2005) studied the combined effects of irradiation
(2 and 5 kGy) and high pressure (33, 400, and 700 MPa, 3 min, 24 and 55°C) pretreatments on
hydration behavior of navy beans. Under moderate pressure (33 MPa), treated beans resulted in a
higher initial moisture uptake (0.59–1.02 kg/kg drymass) and a reduced loss of soluble materials.
The final moisture content after soaking (3 h) was the highest in irradiated beans (5 kGy), followed
by HPP (33 MPa, 3 min, 55°C). However, elevated pressure (400 and 700 MPa) did not signifi-
cantly influence the water uptake due to complete damage of the microstructure, resulting in loss
of soluble solids and lower saturation moisture content (Zhang et al., 2004). Ahromrit et al. (2006)
explored the use of high pressures (up to 600 MPa) to enhance the water uptake during soaking
of glutinous rice. Zhang et al. (2004) studied the effect of high-pressure treatment (300–500 MPa,
0–380 min, 20°C) on water uptake capacity of soybeans and changes in their microstructure. The
NMR analysis revealed that, in high-pressure soaked soybeans, the mobility of water was more
restricted and its distribution was much more uniform as compared to control. Nunez-Mancilla
et al. (2014) indicated that the rehydration ratio, as well as the water holding capacity (WHC)
of strawberry slices, increased as treatment pressure was increased. The maximum WHC was
observed at 500 MPa, while the minimum was noted at 200 MPa. Baier et al. (2015) demonstrated
that rehydration rate of whole peas subjected to high-pressure treatment (400 MPa, 10 min, 40°C)
was higher as compared to conventional thermal treatment (80°C, 10 min). Shang et al. (2015)
demonstrated that at 400 MPa the water retention capacity of sea bass skeletal muscle was higher
as compared to control. Hulle and Rao (2016) indicated that the rehydration ratio of aloe vera
cubes was higher the for the high-pressure pre-treated samples. Ahmed et al. (2016a) indicated
that WHC and water solubility index of high-pressure-treated (300–600 MPa) Thai jasmine rice
flour samples increased linearly with high pressure. Ahmed et al. (2016b) also studied the impact
of high-pressure pre-treatment (400–600 MPa/10 min) on functional, rheological, and pasting
properties of lentil starch dispersions. WHC of the starch increased linearly with pressure due to
increase in damaged starch. High-pressure treatment resulted in the increase in the size of starch
granules as well as resistant starch content.
3.6 EFFECt OF hIGh prESSUrE ON FrEEZING
High-pressure freezing (HPF) is a novel freezing technique that has the potential to manage the
formation and distribution of ice crystals and thus improve the quality of frozen foods (Cheng et al.,
2017a). Few reports are available on the pressure shift freezing (PSF) of tofu (Kanda et al., 1992),
oil-in-water emulsions (Levy et al., 2000), gelatin gels (Chevalier et al., 2000a), potato (Knorr
et al., 1998), eggplant (Otero et al., 1998), peach and mango (Otero et al., 2000), pork (Martino
et al., 1998), and fish (Chevalier et al., 2000b, 2001). These reports indicated that pressure-treated
samples had better preserved microstructure as compared to conventional freezing methods. Zhu
et al. (2003) indicated that pressure freezing produced a large amount of fine and regular intracel-
lular ice crystals, which were homogeneously distributed.
Bulut (2014) indicated that a 3.8-log reduction in number of E. coli was observed after a 30 s
pressure treatment at −3°C. Cheng et al. (2017b) indicated that PSF (200 MPa, 20°C, 30 min) could
reduce the denaturation of frozen natural actomysin at a pressure of 300 MPa. Volkert et al. (2012)
observed that a slight denaturation (about 3%) in milk protein treated by HPP (300 MPa, −16°C)
led to a change in the sensory quality. Choi et al. (2016) demonstrated that L* and b* values of
PSF-treated pork increased with increasing pressure levels up to 150 MPa (Figure 3.7a and c).
Meanwhile, PSF did not affect the redness (a*) of pork and resulted in similar a* values among
all samples (Figure 3.7b). On the other hand, the visual appearance of the pork indicated that the
color of PSF-treated meat changed following the 100 MPa treatment (Figure 3.7d), which was due
to oxidation of myoglobin in pork. Hong and Choi (2016) compared the effects of high-pressure
sub-zero temperature (HPST) and PSF on the quality characteristics of abalone slices. HPST caused
high drip loss relative to the control, whereas drip generation was effectively prevented by PSF at
150 MPa.
80
60
L* 40
20
(a) 0
20
15
a*
10
(b) 0
12
9
b*
(c) 0
(d)
Figure 3.7 effect of PsF on Cie color parameters (a–c) and visual appearance (d) of pork meat. Vertical bars
indicate standard error (n = 3). Means with different letters are significantly different (p < 0.05).
(From Choi, M.J. et al., Lebensm. Wiss. Technol., 67, 194–199, 2016. With permission.)
3.7 EFFECt OF hIGh prESSUrE ON thaWING
Traditional thawing methods are usually time consuming and there are higher chances for the
quality deterioration of frozen foods (Schubring et al., 2003). High-pressure thawing is regarded
as an efficient tool in the food industry for improving the kinetics of water-ice phase transition
and minimizing the changes in quality attributes and microbial growth of frozen foods. Makita
(1992) reported that the time required for high-pressure thawing is one third of the conventional
methods. Takai et al. (1991) indicated that high-pressure thawing (5°C) of tuna meat and surimi
blocks required less time than thawing carried out at ambient conditions. Schubring et al. (2003)
indicated that the water loss was reduced when thawing was assisted by high pressure compared to
thawing at atmospheric pressure.
Yoshioka et al. (1999) indicated that samples subjected to 100–300 MPa for 10 min had quality
attributes similar to fresh ones, which were characterized by lower breaking stress and elasticity as
compared to the sample thawed under running water (15°C–17°C). Zhu et al. (2014) compared the
thawing characteristics of an agar gel and salmon muscle in high-pressure thawing (100, 150, and
200 MPa) and water immersion thawing (WIT), wherein HPP resulted in depression of ice melting
temperature, thereby enhancing the process of thawing. For frozen agar gel, the total thawing time
was 68.7 ± 4.3, 50.3 ± 2.7, 36.4 ± 2.2, and 30.8 ± 1.8 min for WIT and high-pressure thawing at
100, 150, and 200 MPa, respectively. For frozen salmon muscle, the thawing time was 58.9 ± 2.8,
41.8 ± 4.7, 37.2 ± 2.6, and 33.8 ± 1.9 min for WIT and high-pressure thawing at 100, 150, and
200 MPa, respectively. Shiba and Furushita (2017) demonstrated that pressure-assisted thawing
using a thermal buffer zone achieved around 6-log reduction of microbial inactivation.
3.8 EFFECt OF hIGh prESSUrE ON FrYING
Deep-fat frying has been a widely used food processing operation since ancient times and
consists basically of immersion of food pieces in hot vegetable oil. During frying, which involves
simultaneous heat and mass transfer operations, water evaporates from the food into the frying
medium (oil), which in turn penetrates inside the product (Saguy and Dana, 2003). Despite the
appealing characteristics of crispiness, the relatively high oil content (in the range of 33%–38%;
Moreira et al., 1999; Miranda and Aguilera, 2006) is of great concern due to its health as well
as economic impacts. Over the last few decades, several research studies have been carried out
on different approaches to reduce oil uptake, and different mechanisms have been proposed to
explain the phenomenon. However, oil uptake is a complex process, and the oil content in crisps is
relatively high. Hence, further studies are required to understand the various parameters involved
in the process (Dana and Saguy, 2006; Ziaiifar et al., 2008). Al-Khusaibi and Niranjan (2012)
also indicated that the blanching of potato slices (65°C, 5 min) prior to high-pressure treatment
(200–800 MPa, 5 min, 25°C) resulted in a marginal decrease in the oil uptake. However, use of high
pressure prior to frying without blanching resulted in higher absorption of oil. HPP pre-treatment
was found to be helpful in decreasing the frying time required to achieve final desired moisture
content. Albertos et al. (2016) indicated that high-pressure pre-treatment improved the crispiness
and antioxidant properties of vacuum-fried carrot chips compared to control.
3.9 BENEFItS aND LIMItatIONS OF hpp
Though HPP has numerous advantages, there are certain limitations to its application and
feasibility. In general, HPP causes no significant losses of functional compounds in foods but it is
not considered to be a good option for foods that have lower water activity like dry fruits, spices,
powders, and food materials packaged in rigid containers (tin cans, glass bottles). There are also
no published reports available on the toxicity of high-pressure-processed foods. Denys et al. (2000)
indicated that temperature history of a product under pressure is also essential for the optimization
and design of industrial processes. Moreover, research is needed to evaluate pressure uniformity
within a larger pressure volume. A combined treatment of high pressure and temperature is fre-
quently considered to be the most appropriate (Farr, 1990; Patterson et al., 1995). Bacterial spores
are highly pressure resistant, and pressures exceeding 1200 MPa may be needed for their elimination
from foods—this represent the biggest challenge for food safety (Knorr, 1995). Process temperature
in the range 90°C–121°C in conjunction with pressures (>800 MPa) has been used to inactivate
endospore-forming bacteria such as Clostridium botulinum. Thus, low-acid food (pH > 4.6) steril-
ization will most probably rely on a combination of high pressure and other processing treatments
(Cheftel, 1992).
3.10 CONCLUDING rEMarKS
High pressure-processed foods have gained a tremendous advantage over foods processed by
other means, such that they have the potential to be marketed as value-added food products due
to the retention of sensory and nutritional qualities similar to those of fresh ones. HPP is gaining
popularity in the food industry, not only because of its food preservation capacity but also because
of its tremendous potential to achieve interesting functional effects. As a new and novel technology,
HPP has realized success in the food industry that may achieve food safety standards as well as
meet the demand for fresh-tasting minimally processed foods. For years to come, HPP is likely to
be used commercially on a wider scale as it offers a natural alternative for the processing of a wide
range of different food products.
rEFErENCES
Ade-Omowaye, B.I., Rastogi, N.K., Angersbach, A., and Knorr, D. (2001). Effect of high pressure or high
electrical field pulse pretreatment on dehydration characteristics of paprika. Innov Food Sci Emerg
Technol. 2: 1–7.
Ade-Omowaye, B.I.O., Rastogi, N.K., Angersbach, A., and Knorr, D. (2003). Combined effects of pulsed
electric field pre-treatment and partial osmotic dehydration on air drying behaviour of red bell pepper.
J Food Eng. 60: 89–98.
Ahmed, J., Mulla, M.Z., Arfat, Y.A., and Kumar, V. (2016a). Effects of high-pressure treatment on functional,
rheological, thermal and structural properties of Thai jasmine rice flour dispersion. J Food Process
Preserv. 41: 1–12.
Ahmed, J., Thomas, L., Taher, A., and Joseph, A. (2016b). Impact of high pressure treatment on functional,
rheological, pasting and structural properties of lentil starch dispersions. Carbohydr Polym. 152:
639–647.
Ahromrit, A., Ledward, D.A., and Niranjan, K. (2006). High pressure induced water uptake characteristics of
Thai glutinous rice. J Food Eng. 72: 225–233.
Albertos, I., Martin-Diana, A.B., Sanz, M.A., Barat, J.M., Diez, A.M., Jaime, I., and Rico, D. (2016). Effect of
high pressure processing or freezing technologies as pretreatment in vacuum fried carrot snacks. Innov
Food Sci Emerg Technol. 33: 115–122.
Al-Khusaibi, M.K., and Niranjan, K. (2012). The impact of blanching and high-pressure pretreatments on oil
uptake of fried potato slices. Food Bioprocess Technol. 5: 2392–2400.
Al-Khuseibi, M.K., Sablani, S.S., and Perera, C.O. (2005). Comparison of water blanching and high hydro-
static pressure effects on drying kinetics and quality of potato. Drying Technol. 23: 2449–2461.
Altuner, E.M., Ceter, T., and Alpas, H. (2012). High hydrostatic pressure processing: A method having high
success potential in pollen protein extraction. High Press Res. 32: 291–298.
Altuner, E.M., Ceter, T., and Alpas, H. (2014). Effect of high hydrostatic pressure on the profile of proteins
extracted from Betula pendula pollens. High Press Res. 34: 470–481.
Amami, E., Vorobiev, E., and Kechaou, N. (2006).Modelling of mass transfer during osmotic dehydration of
apple tissue pre-treated by pulsed electric field. Lebensm Wiss Technol. 39: 1014–1021.
Azuara, E., Garcia, H.S., and Beristain, C.I. (1996). Effect of centrifugal force on osmotic dehydration of
potatoes and apples. Food Res Int. 29: 195–199.
Baier, A.K., Bubler, S., and Knorr, D. (2015). Potential of high isostatic pressure and pulsed electric fields to
improve mass transport in pea tissue. Food Res Int. 76: 66–73.
Balasubramaniam, V.M. (2003). High pressure food preservation. In: Encyclopedia of Agriculture, Food and
Biological Engineering, pp. 490–496. Heldman, D., Ed., Marcel Dekker, New York.
Balasubramaniam, V.M., and Farkas, D. (2008). High pressure food processing. Food Sci Technol Int. 14:
413–418.
Balci, A.T., and Wilbey, R.A. (1999). High pressure processing of milk: The first 100 years in the development
of new technology. Int J Dairy Technol. 52: 149–155.
Bi, H.M., Shou, Q., Zhang, C., Jiao, L., and Chang, Z.W. (2009). High hydrostatic pressure extraction of sali-
droside from Rhodiola sachalinensis. J Food Process Eng. 32: 53–63.
Briones-Labarca, V., Plaza-Morales, M., Giovagnoli-Vicuna, C., and Jamett, F. (2015). High hydrostatic pres-
sure and ultrasound extractions of antioxidant compounds, sulforaphane and fatty acids from Chilean
papaya (Vasconcellea pubescens) seeds: Effects of extraction conditions and methods. Lebensm Wiss
Technol. 60: 525–534.
Bulut, S. (2014). Inactivation of Escherichia coli in milk by high pressure processing at low and subzero tem-
peratures. High Pressure Res. 34: 439–446.
Casquete, R., Castro, S.M., Villalobos, M.C., Serradilla, M.J., Queiros, R.P., Saraiva, J.A., Cordoba, M.G.,
and Teixeira, P. (2014). High pressure extraction of phenolic compounds from citrus peels. High Press
Res. 34: 447–451.
Cheftel, J.C. (1992). Effects of high hydrostatic pressure on food constituents: An overview. In: High Pressure
and Biotechnology, pp. 195–209. Balny, C., Hayashi, R., Heremans, K., and Masson, P., Eds., INSER
My John Libbey Eurotext, Paris, France.
Chemat, F., Fabiano-Tixier, A.S., Vian, M.A., Allaf, T., and Vorobiev, E. (2015). Solvent-free extraction of
food and natural products. Trends Anal Chem; TrAC. 71: 157–168.
Chemat, F., Vian, M.A., and Cravotto, G. (2012). Green extraction of natural products: Concept and principles.
Int J Mol Sci. 13: 8615–8627.
Chen, R., Meng, F., Zhang, S., and Liu, Z. (2009). Effects of ultra high pressure extraction conditions on yields
and antioxidant activity of ginsenoside from ginseng. Sep Purif Technol. 66: 340–346.
Cheng, L., Sun, D.W., Zhu, Z., and Zhang, Z. (2017a). Emerging techniques for assisting and accelerating food
freezing processes: A review of recent research progresses. Crit Rev Food Sci Nutr. 57: 769–781.
Cheng, L., Sun, D.W., Zhu, Z., and Zhang, Z. (2017b). Effects of high pressure freezing (HPF) on denaturation
of natural actomyosin extracted from prawn (Metapenaeus ensis). Food Chem. 229: 252–259.
Chevalier, D., Le Bail, A., and Ghoul, M. (2000a). Freezing and ice crystals formed in a cylindrical food
model: Part 11, the comparison between freezing at atmospheric pressure and pressure shift freezing.
J Food Eng. 46: 287–293.
Chevalier, D., Sentissi, M., Havet, M., and Le Bail, A. (2000b). Comparison of air-blast and pressure shift
freezing on Norway lobster quality. J Food Sci. 65: 329–333.
Chevalier, D., Sequeira-Munoz, A., Le Bail, A., Simpson, B.K., and Ghoul, M. (2001). Effect of freezing
conditions and storage on ice crystal and drip volume in turbot (Scophthalmus maximus), evaluation of
pressure shift freezing vs. air-blast freezing. Innov Food Sci Emerg Technol. 1: 193–201.
Choi, M.J., Min, S.G., and Hong, G.P. (2016). Effects of pressure-shift freezing conditions on the quality char-
acteristics and histological changes of pork. Lebensm Wiss Technol. 67: 194–199.
Corrales, M., Garc, A.F., Butz, P., and Tauscher, B. (2009). Extraction of anthocyanins from grape skins
assisted by high hydrostatic pressure. J Food Eng. 90: 415–421.
Cullen, P.J., Tiwari, B.K., and Valdramidis, V.P. (2012). Status and trends of novel thermal and non-thermal
technologies for fluid foods. In: Novel Thermal and Non-thermal Technologies for Fluid Foods, pp. 1–6.
Valdramidis, V.P., Ed., Academic Press, San Diego, CA.
Dana, D., and Saguy, I.S. (2006). Review: Mechanism of oil uptake during deep-fat frying and the surfactant
effect-theory and myth. Adv Colloid Interface Sci. 128–130: 267–272.
Denys, S., Van Loey, A.M., and Hendrickx, M.E. (2000). Modeling conductive heat transfer during high-
pressure thawing processes: Determination of latent heat as a function of pressure. Biotechnol Prog.
16: 447–455.
Dermesonlouoglou, E.K., Andreou, V., Alexandrakis, Z., Katsaros, G.J., Giannakourou, M.C., and Taoukis,
P.S. (2017). The hurdle effect of osmotic pretreatment and high-pressure cold pasteurisation on the shelf-
life extension of fresh-cut tomatoes. Int J Food Sci Technol. 52: 916–926.
Dornenburg, H., and Knorr, D. (1993). Cellular permeabilisation of cultured plant tissue by high electric field
pulse and ultra high pressure for recovery of secondary metabolites. J Food Biotechnol. 7: 35–48.
Duvetter, T., Fraeye, I., Van Hoang, T., Buggenhout, S.V., Verlent, I., Smout, C., Loey, A.V., and Hendrickx,
M. (2005). Effect of pectin methylesterase infusion methods and processing techniques on strawberry
firmness. J Food Sci. 70: S383–S388.
Eshtiaghi, M.N., Stute, R., and Knorr, D. (1994). High pressure and freezing pretreatment effects on drying,
rehydration, texture and color of green beans, carrots and potatoes. J Food Sci. 59: 1168–1170.
Farr, D. (1990). High-pressure technology in food industry. Trends Food Sci Technol. 1: 14–16.
Ferrari, G., Maresca, P., and Ciccarone, R. (2010). The application of high hydrostatic pressure for the stabili-
zation of functional foods: Pomegranate juice. J Food Eng. 100: 245–253.
Fito, P., Chiralt, N., Betoret, N., Gras, M., Chafer, M., Martinez-Monzo, J., Andres, A., and Vidal, D. (2001).
Vacuum impregnation and osmotic dehydration in matrix engineering application in functional fresh
food development. J Food Eng. 49: 175–183.
Fraeye, I., Knockaert, G., Van Buggenhout, S., Duvetter, T., Hendrickx, M., and Loey, A.V. (2010). Enzyme
infusion prior to thermal/high pressure processing of strawberries: Mechanistic insight into firmness
evolution. Innov Food Sci Emerg Technol. 11: 23–31.
Galazka, V.B., and Ledward, D.A. (1995). Development in high pressure processing. Food Technol Int Eur.
12: 123–125.
George, J.M., and Rastogi, N.K. (2016). High pressure processing for food fermentation. In: Novel Food
Fermentation Technologies, pp. 57–83. Ojha, K.S., and Tiwari, B.K., Eds., Springer International
Publishing, Switzerland.
George, J.M., Selvan, T.S., and Rastogi, N.K. (2016). High pressure assisted infusion of bioactive compounds
in apple slices. Innov Food Sci Emerg Technol. 33: 100–107.
Gould, G.W. (1995). The microbe as a high-pressure target. In: High Pressure Processing of Foods, pp. 27–35.
Ledward, D.A., Johnston, D.E., Earnshaw, R.G., and Hasting, A.P.M., Eds., Nottingham University
Press, Nottingham, UK.
Grossi, A., Soltoft-Jensen, J., Knudsen, J.C., and Christensen, M. (2011). Synergistic cooperation of high pres-
sure and carrot dietary fibre on texture and colour of pork sausages. Meat Sci. 89: 195–201.
Guo, X., Han, D., Xi, H., Rao, L., Liao, X., Hu, X., and Wu, J. (2012). Extraction of pectin from navel orange
peel assisted by ultra-high pressure, microwave or traditional heating: A comparison. Carbohydr Polym.
88: 441–448.
Hayashi, R. (1990). Application of high pressure to processing and preservation: Philosophy and development.
In: Engineering and Food, pp. 815–826. Spiess, W.E.L., and Schubert, H., Eds., Elsevier, London, UK.
Hite, B.H. (1899). The effect of pressure in the preservation of milk. Wash Va Univ Agric Exp StnBull. 58:
15–35.
Hite, B.H., Giddings, N.J., and Weakly, C.E. (1914). The effects of pressure on certain microorganisms
encountered in the preservation of fruits and vegetables. Wash Va Univ Agric Exp Stn Bull. 146: 1–67.
Hogan, E., Kelly, A., and Sun, D.W. (2005). High pressure processing of foods: An overview. In: Emerging
Technologies for Food Processing, pp. 3–32. Sun, D.W., Ed., Elsevier Academic Press, London, UK.
Hong, G.P., and Choi, M.J. (2016). Comparison of the quality characteristics of abalone processed by high-
pressure sub-zero temperature and pressure-shift freezing. Innov Food Sci Emerg Technol. 33: 19–25.
Hu, W., Guo, T., Jiang, W.J., Dong, G.L., Chen, D.W., Yang, S.L., and Li, H.R. (2015). Effects of ultra high
pressure extraction on yield and antioxidant activity of chlorogenic acid and cynaroside extracted from
flower buds of Lonicera japonica. Chin J Nat Med. 13: 445–453.
Hugas, M., Garriga, M., and Monfort, J.M. (2002). New mild technologies in meat processing: High pressure
as a model technology. Meat Sci. 62: 359–371.
Hulle, N.R.S., and Rao, P.S. (2016). Effect of high pressure pretreatments on structural and dehydration char-
acteristics of aloe vera (Aloe barbadensis Miller) cubes. Drying Technol. 34: 105–118.
Jangam, S.V. (2011). An overview of recent developments and some R&D challenges related to drying of
foods. Drying Technol. 29: 1343–1357.
Janowicz, M., and Lenart, A. (2015). Selected physical properties of convection dried apples after HHP treat-
ment. Lebensm Wiss Technol. 63: 828–836.
Jung, S. (2016). Applications and opportunities for pressure-assisted extraction. In: High Pressure Processing
of Foods, pp. 173–191. Balasubramaniam, V.M., Barbosa-Canovas, G.V., Lelieveld, H.L.M., Eds.,
Springer, New York.
Kanda, Y., Aoki, M., and Kosugi, T. (1992). Freezing of tofu (soyabean curd) by pressure shift freezing and its
structure. J Jpn Soc Nutr Food Sci. 39: 608–614.
Kim, S., Lee, Y.R., Noh, Y.H., Lee, J., and Jeong, H.S. (2015). Effects of high hydrostatic pressure treatment
on the enhancement of functional components of germinated rough rice (Oryza sativa L.). Food Chem.
166: 86–92.
Kim, M.B., Park, J.E., Woo, S.W., Lim, S.B., and Hwang, J.K. (2014). Optimization of high hydrostatic
pressure process for the extraction of kirenol from Siegesbeckia orientalis l. using response surface
methodology. Food Sci Biotechnol. 23: 731–738.
Knorr, D. (1995). Hydrostatic pressure treatment of food: Microbiology. In: New Methods for Food
Preservation, pp. 159–175. Gould, G.W., Ed., Blackie Academic and Professional, London, UK.
Knorr, D., Schlueter, O., and Heinz, V. (1998). Impact of high hydrostatic pressure on phase transitions of
foods. J Food Technol. 52: 42–45.
Kowalski, S.J., Szadzinska, J., and Pawłowski, A. (2015). Ultrasonic assisted osmotic dehydration of car-
rot followed by convective drying with continuous and intermittent heating. Drying Technol. 33:
1570–1580.
Lakshmanana, R., Parkinson, J.A., and Piggott, J.R. (2007). High-pressure processing and water-holding
capacity of fresh and cold-smoked salmon (Salmo salar). Lebensm Wiss Technol. 40: 544–551.
Lee, H.S., Lee, H.J., Yu, H.J., Judo, W., Kim, Y., Kim, C.T., Kim, C.J., Cho, Y.J., Kim, N., Choi, S.Y., and
Suh, H.J. (2011). A comparison between high hydrostatic pressure extraction and heat extraction of
ginsenosides from ginseng (Panax ginseng CA Meyer). J Sci Food Agric. 91: 1466–1473.
Leng, X.J., Zhang, L., Huang, M., Xu, X.L., and Zhou, G.H. (2013). Mass transfer dynamics during high
pressure brining of chicken breast. J Food Eng. 118: 296–301.
Levy, J., Dumay, E., Kolodziejczyk, E., and Cheftel, J.C. (2000). Kinetics of freezing of an oil-in-water
emulsion by pressure release. Impact on ice crystals and oil droplets. High Press Res. 19: 183–189.
Li, J., Sun, W., Ramaswamy, H.S., Yu, Y., Zhu, S., and Wang, J., and Li, H. (2016). High pressure extraction
of astaxanthin from shrimp waste (Penaeus vannamei boone): Effect on yield and antioxidant activity.
J Food Process Eng. doi:10.1111/jfpe.12353.
Mahadevan, S., Nitin, D., Salvi, D., and Karwe, M.V. (2014). High-pressure enhanced infusion: Influence of
process parameters. J Food Process Eng. 38: 601–612.
Makita, T. (1992). Application of high pressure and thermophysical properties of water to biotechnology.
Fluid Phase Equilibr. 76: 87–95.
Martino, M.N., Otero, L., Sanz, P.D., and Zaritzky, N.E. (1998). Size and location of ice crystals in pork frozen
by high-pressure-assisted freezing as compared to classical methods. Meat Sci. 50: 303–313.
Miles, S., Braxton, D.S., and Frewer, L.J. (1999). Public perceptions about microbiological hazards in food.
Br Food J. 101: 744–762.
Miranda, M.L., and Aguilera, J.M. (2006). Structure and texture properties of fried potato products. Food Rev
Int. 22: 173–201.
Moreira, R.G., Castell-Perez, M.E., and Barrufet, M.A. (1999). Deep Fat Frying Fundamentals and
Applications. Springer, London, UK.
Mothibe, K.J., Zhang, M., Nsor-Atindana, J., and Wang, Y.C. (2011). Use of ultrasound pretreatment in drying
of fruits: Drying rates, quality attributes, and shelf life extension. Drying Technol. 29: 1611–1621.
Nunez-Mancilla, Y., Perez-Won, M., Vega-Galvez, A., Arias, V., Tabilo-Munizaga, G., Briones-Labarca, V.,
Lemus-Mondaca, R., and Scala, K.D. (2011). Modeling mass transfer during osmotic dehydration of
strawberries under high hydrostatic pressure conditions. Innov Food Sci Emerg Technol. 12: 338–343.
Nunez-Mancilla, Y., Vega-Galvez, A., Perez-Won, M., Zura, L., Garcia-Segovia, P., and Scala, K.D. (2014).
Effect of osmotic dehydration under high hydrostatic pressure on microstructure, functional properties
and bioactive compounds of strawberry (Fragaria Vesca). Food Bioprocess Technol. 7: 516–524.
Ohlsson, T., and Bengtsson, N. (2002). Minimal Processing Technologies in the Food Industry. Woodhead
Publishing Ltd, Cambridge, UK.
Oliveira, M.M., Tribst, A.A.L., Junior, C.L., and Oliveira, R.A. (2015). Effects of high pressure processing on
cocoyam, Peruvian carrot, and sweet potato: Changes in microstructure, physical characteristics, starch,
and drying rate. Innov Food Sci Emerg Technol. 31: 45–53.
Olsson, S. (1995). Production equipment for commercial use. In: High Pressure Processing of Foods, pp.
167–180. Ledward, D.A., Johnston, D.E., Earnshaw, R.G., and Hastings, A.P.M., Eds., Nottingham
University Press, Nottingham, UK.
Otero, L., Martino, M., Zaritzky, N.E., Solas, M., and Sanz, P.D. (2000). Preservation of microstructure in
peach and mango during high pressure-shift freezing. J Food Sci. 65: 466–470.
Otero, L., Solas, M.T., Sanz, P.D., de Elvira, C., and Carrasco, J.A. (1998). Contrasting effects of high pressure
assisted freezing and conventional air freezing on eggplant tissue microstructure. Eur Food Res Technol.
206: 338–342.
Paini, M., Casazza, A.A., Aliakbarian, B., Perego, P., Binello, A., and Cravotto, G. (2016). Influence of ethanol/
water ratio in ultrasound and high‐pressure/high‐temperature phenolic compound extraction from agri‐
food waste. Int J Food Sci Technol. 51: 349–358.
Pan, Y.K., Zhao, L.J., Zhang, Y., Chen, G., and Mujumdar, A.S. (2003). Osmotic dehydration pretreatment in
drying of fruits and vegetables. Drying Technol. 21: 1101–1114.
Park, C.Y., Kim, S., Lee, D., Park, D.J., and Imm, J.Y. (2016). Enzyme and high pressure assisted extraction
of tricin from rice hull and biological activities of rice hull extract. Food Sci Biotechnol. 25: 159–164.
Passos, J.J., De Sousa, F.B., Mundim, I.M., Bonfim, B.R., Melo, R., Viana, A.F., Stolz, E.D., Borsoi, M., Rates,
S.M.K., and Sinisterra, R.D. (2013). Double continuous injection preparation method of cyclodextrin
inclusion compounds by spray drying. Chem Eng J. 228: 345–351.
Patterson, M.F., Quinn, M., Simpson, R., and Gilmour, A. (1995). Effect of high pressure on vegetable patho-
gens. In: High Pressure Processing of Foods, pp. 47–63. Ledward, D.A., Johnston, D.E., Earnshaw,
R.G., Hasting, A.P.M., Eds., Nottingham University Press, Nottingham, UK.
Pauling, L. (1964). College Chemistry: An Introductory Textbook of General Chemistry. Freeman and
Company, San Francisco, CA.
Perez-Lopez, E., Mateos-Aparicio, I., and Ruperez, P. (2016). Okara treated with high hydrostatic pressure
assisted by Ultraflo® L: Effect on solubility of dietary fibre. Innov Food Sci Emerg Technol. 33: 32–37.
Perez-Won, M., Lemus-Mondaca, R., Tabilo-Munizaga, G., Pizarro, S., Noma, S., Igura, N., and Shimoda, M.
(2016). Modelling of red abalone (Haliotis rufescens) slices drying process: Effect of osmotic dehydra-
tion under high pressure as a pretreatment. Innov Food Sci Emerg Technol. 34: 127–134.
Plaza, L., Colina, C., de Ancos, B., Sanchez-Moreno, C., and Cano, M.P. (2012). Influence of ripening and
astringency on carotenoid content of high-pressure treated persimmon fruit (Diospyros kaki L.). Food
Chem. 130: 591–597.
Plaza, L., Sanchez-Moreno, C., de Ancos, B., Elez-Martinez, P., Martın-Belloso, O., and Cano, M.P. (2011).
Carotenoid and flavanone content during refrigerated storage of orange juice processed by high-pressure,
pulsed electric fields and low pasteurization. Lebensm Wiss Technol. 44: 834–849.
Prasad, N.K., Hao, J., Shi, J., Liu, T., Li, J., Wei, X., Qiu, S., Xue, S., and Jiang, Y. (2009b). Antioxidant and
anticancer activities of high pressure-assisted extract of longan (Dimocarpus longan Lour.) fruit pericarp.
Innov Food Sci Emerg Technol. 10: 413–419.
Prasad, K.N., Yang, B., Zhao, M., Wang, B.S., Chen, F., and Jiang, Y. (2009a). Effects of high-pressure treat-
ment on the extraction yield, phenolic content and antioxidant activity of litchi (Litchi chinensis Sonn.)
fruit pericarp. Int J Food Sci Technol. 44: 960–966.
Raikham, C., Prachayawarakorn, S., Nathakaranakule, A., and Soponronnarit, S. (2015). Influences of pre-
treatments and drying process including fluidized bed puffing on quality attributes and microstructural
changes of banana slices. Drying Technol. 33: 915–925.
Ramaswamy, R., Balasubramaniam, V.M., and Sastry, S.K. (2005). Effect of high pressure and irradiation
treatments on hydration characteristics of navy beans. Int J Food Eng. 1: 1–17.
Rastogi, N.K. (2011). Opportunities and challenges in application of ultrasound in food processing. Crit Rev
Food Sci Nutr. 51: 705–722.
Rastogi, N.K., Angersbach, A., Niranjan, K., and Knorr, D. (2000). Rehydration kinetics of high pressure
treated and osmotically dehydrated pineapple. J Food Sci. 65: 838–841.
Rastogi, N.K., Nguyen, L.T., and Balasubramaniam, V.M. (2008). Effect of pre-treatments on carrot texture
after thermal and pressure-assisted thermal processing. J Food Eng. 88: 541–547.
Rastogi, N.K., Nguyen, L.T., Jiang, B., and Balasubramaniam, V.M. (2010). Improvement in texture of
pressure-assisted thermally processed carrots by combined pretreatment using response surface meth-
odology. Food Bioprocess Technol. 3: 762–771.
Rastogi, N.K., and Niranjan, K. (1998). Enhanced mass transfer during osmotic dehydration of high pressure
treated pineapple. J Food Sci. 63: 508–511.
Rastogi, N.K., and Raghavarao, K.S.M.S. (1996). Kinetics of osmotic dehydration under vacuum. Lebensm
Wiss Technol. 29: 669–672.
Rastogi, N.K., Raghavarao, K.S.M.S., Balasubramaniam, V.M., Niranjan, K., and Knorr, D. (2007).
Opportunities and challenges in high pressure processing of foods. Crit Rev Food Sci Nutr. 47: 69–112.
Rastogi, N.K., Subramanian, R., and Raghavarao, K.S.M.S. (1994). Application of high-pressure technology
in food processing. Indian Food Ind. 13: 30–34.
Rendueles, E., Omer, M.K., Alvseike, O., Alonso-Calleja, C., Capita, R., and Prieto, M. (2011). Microbiological
food safety assessment of high hydrostatic pressure processing: A review. Lebensm Wiss Technol. 44:
1251–1260.
Rodrigues, D., Freitas, A.C., Queiros, R., Rocha-Santos, T.A.P., Saraiva, J.A., Gomes, A.M.P., and Duarte,
A.C. (2016). Bioactive polysaccharides extracts from Sargassum muticum by high hydrostatic pressure.
J Food Process Eng. 41:1–12. doi:10.1111/jfpp.12977.
Rombaut, N., Tixier, A.S., Bily, A., and Chemat, F. (2014). Green extraction processes of natural products as
tools for biorefinery. Biofuel Bioprod Bior. 8: 530–544.
Saguy, I.S., and Dana, D. (2003). Integrated approach to deep fat frying: Engineering, nutrition, health and
consumer aspects. J Food Eng. 56: 143–152.
Sale, A.J., Gould, G.W., and Hamilton, W.A. (1970). Inactivation of bacterial spores by hydrostatic pressure.
J Gen Microbiol. 60: 323–334.
Santos, M.C., Salvador, A.C., Domingues, F.M., Cruz, J.M., and Saraiva, J.A. (2013). Use of high hydrostatic
pressure to increase the content of xanthohumol in beer wort. Innov Food Sci Emerg Technol. 13: 13–22.
Schubring, R., Meyer, C., Schlüter, O., Boguslawski, S., and Knorr, D. (2003). Impact of high pressure assisted
thawing on the quality of fillets from various fish species. Innov Food Sci Emerg Technol. 4: 257–267.
Seo, Y.C., Choi, W.Y., Kim, J.S., Yoon, C.S., Lim, H.W., and Cho, J.S. (2011). Effect of ultrahigh pressure
processing on immuno-modulatory activities of the fruits of Rubus coreanus miquel. Innov Food Sci
Emerg Technol. 12: 207–215.
Shang, X., Liu, A., Zheng, J., Wang, P., and Yin, S. (2015). High pressure processing and water holding capac-
ity of sea bass skeletal muscle. J Aquat Food Prod T. 24: 740–751.
Shiba, T., and Furushita, M. (2017). Pressure-induced inactivation of bacteria through pressure-assisted
thawing using a thermal buffer zone. Innov Food Sci Emerg Technol. 39: 19–24.
Shouqin, Z., Ruizhan, C., and Changzheng, W. (2007). Experiment study on ultrahigh pressure extraction of
ginsenosides. J Food Eng. 79: 1–5.
Sopanangkul, A., Ledward, D.A., and Niranjan, K. (2002). Mass transfer during sucrose infusion into potatoes
under high pressure. J Food Sci. 67: 2217–2220.
Strati, I.F., Gogou, E., and Oreopoulou, V. (2015). Enzyme and high pressure assisted extraction of carotenoids
from tomato waste. Food Bioprod Process. 94: 668–674.
Takai, R., Kozhima, T.T., and Suzuki, T. (1991). Low temperature thawing by using high-pressure. In: Les
Actes du XVIIIe Congres International du Froid, Montreal, Canada, pp. 1951–1955.
Thakur, B.R., and Nelson, P.E. (1998). High pressure processing and preservation of foods. Food Rev Int. 14:
427–447.
Tola, Y.B., and Ramaswamy, H.S. (2013). Evaluation of high pressure treatment (HP) for rapid and uniform
pH reduction in carrots. J Food Eng. 116: 900–909.
Vega-Galvez, A., Uribe, E., Perez, M., Tabilo-Munizaga, G., Vergara, J., Garcia-Segovia, P., Lara, E., and
Scala, K.D. (2011). Effect of high hydrostatic pressure pretreatment on drying kinetics, antioxidant
activity, firmness and microstructure of aloe vera (Aloe barbadensis Miller) gel. Lebensm Wiss Technol.
44: 384–391.
Verma, D., Kaushik, N., and Rao, P.S. (2014). Application of high hydrostatic pressure as a pretreatment for
osmotic dehydration of banana slices (Musa cavendishii) finish-dried by dehumidified air drying. Food
Bioprocess Technol. 7: 1281–1297.
Volkert, M., Puaud, M., Wille, H.J., and Knorr, D. (2012). Effects of high pressure-low temperature treatment
on freezing behavior, sensorial properties and air cell distribution in sugar rich dairy based frozen food
foam and emulsions. Innov Food Sci Emerg Technol. 13: 75–85.
Won, Y.C., Min, S.C., and Lee, D.U. (2015). Accelerated drying and improved color properties of red pepper
by pretreatment of pulsed electric fields. Drying Technol. 33: 926–932.
Xi, J. (2006). Application of high hydrostatic pressure processing of food to extracting lycopene from tomato
paste waste. High Press Res. 26: 33–41.
Yan, H., Zhang, S., Luo, X., Hou, L., and Liang, Q. (2008). HHPE: Improving the stability and antioxidant
activity of bioactive components from Hedyotis diffusa willd. J Biotechnol. 136: 468–469.
Yang, B., Jiang, Y., Wang, R., Zhao, M., and Sun, J. (2009). Ultra-high pressure treatment effects on polysac-
charides and lignins of longan fruit pericarp. Food Chem. 112: 428–431.
Yoshioka, K., Yamada, A., Maki, T., Yoshimoto, C., and Yamamoto, T. (1999). Application of high pressurisa-
tion to fish meat: The ultrastructural changes and nucleotide in frozen carp muscle under high pressure
thawing. In: Advances in High Pressure Bioscience and Biotechnology, pp. 347–369. Ludwig, H., Ed.,
Springer, Berlin, Germany.
Yucel, U, Alpas, H., and Bayindirli, A. (2010). Evaluation of high pressure pretreatment for enhancing the
drying rates of carrot, apple, and green bean. J Food Eng. 98: 266–272.
Zhang, H., Ishida, N., and Isobe, S. (2004). High-pressure hydration treatment for soybean processing. Trans
ASAE. 47: 1151–1158.
Zhang, S., Wang, Z., Wang, T., Zheng, N., Li, M., and Lin, J. (2012). Optimization of central composite design-
response surface methodology in ultra high pressure extraction of Scutellaria baicalensis. J Med Plants
Res. 6: 373–378.
Zhu, S., Bail, A.L., and Ramaswamy, H.S. (2003). Ice crystal formation in pressure shift freezing of Atlantic
salmon (Salmo salar) as compared to classical freezing methods. J Food Process Preserv. 27: 427–444.
Zhu, S., Su, G.M., He, J.S., Ramaswamy, H.S., Le Bail, A., and Yu, Y. (2014). Water phase transition under
pressure and its application in high pressure thawing of agar gel and fish. J Food Eng. 125: 1–6.
Ziaiifar, A.M., Achir, N., Courtois, F., Trezzani, I., and Trystram, G. (2008). Review of mechanisms, condi-
tions, and factors involved in the oil uptake phenomenon during the deep-fat frying process. Int J Food
Sci Technol. 43: 1410–1423.
ChaptEr 4
high-pressure processing of Meat,

Fish, and poultry products
K. Jayathilakan, Khudsia Sultana and M. C. Pandey
CONtENtS
4.1 Introduction ............................................................................................................................ 47

4.2 Principle and Mechanism ....................................................................................................... 48
4.3 High-Pressure Processing of Meat and Poultry Products ...................................................... 49
4.3.1 Impact of HPP on Colour of Meat .............................................................................. 50
4.3.2 Impact of HPP on Flavour .......................................................................................... 52
4.3.3 Impact of HPP on Texture .......................................................................................... 53
4.3.4 Impact of HPP on Lipids ............................................................................................ 53
4.3.5 Impact of HPP on Non-heme Iron .............................................................................. 54
4.3.6 Impact of HPP on Proteins ......................................................................................... 54
4.3.7 Impact of HPP on Thermal Behaviour ....................................................................... 55
4.3.8 Impact of HPP on Water Holding Capacity................................................................ 56
4.4 Effect of HPP on Microorganisms ......................................................................................... 57
4.5 Applications of HPP in Meat, Poultry, and Fish..................................................................... 58
4.6 Combined Treatments with High-Pressure Processing .......................................................... 59
4.7 Summary ................................................................................................................................ 61
References ........................................................................................................................................ 62
4.1 INtrODUCtION
High-pressure processing (HPP) is one of the innovative and successful non-thermal preserva-
tion techniques which can be employed for the development of minimally processed and other
perishable commodities. HPP treats products statically at or above 100 MPa by means of a liquid
transmitter. According to Patterson (2005), it is common to use pressure from 500 to 900 MPa. HPP
has various advantages over other non-thermal technologies. Food can be processed at ambient or
even at lower temperatures. In the transmission of pressure, the processed material experiences
the pressure instantaneously with no gradient, resulting in uniform treatment irrespective of the
size and geometry of the material. High-pressure modifies only non-covalent bonds like hydrogen,
ionic, and hydrophobic bonds and does not affect small molecules such as flavour compounds and
vitamins (Toepfl et al., 2006).
Therefore, HPP leads to less degradation in the overall quality of processed foods as compared
to heat-treated foods. In addition, HPP takes less time and lower energy (Simonin et al., 2012).
47
HPP has been applied as a preservation method to a wide range of meat products such as cured
meats, processed meats, or meats for further processing and ready meals. However, high-pressure
treatment can increase lipid oxidation and induce colour and texture changes in red meat (Yagiz
et al., 2009).
Product shelf life is generally extended due to the inactivation of enzymes and microorganisms
without affecting products sensory and nutritional attributes. HPP can be successfully implemented
in the meat industry by standardising the quality protocols for achieving a product’s shelf stability.
HPP application can be carried out by selecting and standardizing proper pressures with or without
addition of heat to achieve inactivation of microbes or to alter the product attributes to meet the
requirements of consumers. Several factors influence the process of HPP like type of microorgan-
ism, composition of food, pH, and water activity.
HPP produces products with good retention of nutrients and having a fresh-like quality. This
process can be applied for both acid and low acid foods. Some of the drawbacks include the fact that
it is a semicontinuous process, it involves expensive equipment, high pressure may lead to textural
alterations, generation of free radicals leading to lipid oxidation, and its insensitivity towards spores.
Application of HPP has been broadened to food products such as fruit juices and meat products
(Heinz and Buckow, 2010). This technology can offer a high retention of sensory and nutritional
attributes of food products, because the treatment can be performed at room temperature, while
ensuring safety and stability during refrigerated storage.
Consumer demand for fresh-like food products with minimal deterioration of sensory and nutri-
tional properties has initiated research on new emerging non-thermal treatments in the food industry.
In recent years, HPP has gained much attention as a food preservation method which is also known
as cold pasteurization, ultra high-pressure processing, or high hydrostatic processing. It eliminates
spoilage and pathogenic microorganisms (Sun and Norton, 2008) by holding the nutritional and
sensory characteristics of food products (Rastogi et al., 2007). In general, in HPP treatment foods
are submitted to pressure levels ranging from 100 to 900 MPa, although 400–600 MPa pressures
are frequently used for commercial application. The pressure differs from product to product as
non-thermal decontamination technology (Jiménez-Colmenero and Borderias, 2003). The effects
of HPP treatment differ from the conventional thermal processing such as dehydration, irradiation,
and other processing techniques, where these techniques act by inactivating enzymes and reducing
microbial load, thereby preventing spoilage and extending shelf life. However, thermal processing
techniques affect the freshness of the product. HPP, being a non-thermal technique, lengthens shelf
life without affecting its freshness. This technology exhibits uniform effects and is nearly instanta-
neous throughout the food. Thus, it is independent of food geometry and equipment size.
4.2 prINCIpLE aND MEChaNISM
HPP applies very high pressures (up to 6000 times atmospheric pressure) to packages or bulk
liquid and solid foods, in a hydrostatic press. Baskets containing packaged foods are introduced into
steel vessels packed with water, and high pressure is applied (through a piston) to the incompress-
ible water to be transferred to the food uniformly from all surfaces. Since air is more compressible
than water, the presence of air increases the pressurisation time of the food. After completion of
pressurisation, the food has to be kept under required pressure for a particular process time, which
normally may be for several minutes. After this, usually the depressurisation will be a rapid process.
HPP uses an isostatic pressure at room temperature and between 100 and 600 MPa. The pres-
sure chamber is loaded, closed, and degassed and the pressure is transmitted by the pumps through
a liquid, generally water as shown in Figure 4.1. The technology is based on the principle of Le
Chatelier and the isostatic rule, so there is uniform application of pressure instantaneously through
a food material (with or without packaging) independent of its mass, shape, and composition.
HiGH-Pressure ProCessinG oF MeAt, FisH, AnD PouLtry ProDuCts 49
Figure 4.1 operation of an HPP unit. (From http://www.hiperbaric.com.)
In particular, high pressure accelerates reactions involving a change of volume at the molecular
level with partial unfolding of proteins which promote covalent and non-covalent interactions dur-
ing and upon release of pressure, leading to denaturation (Rastogi et al., 2007). HPP causes inacti-
vation of microorganisms and enzymes and changes the rheological properties of food products. In
comparison with other thermal processing techniques, HPP produces foods with fresher taste and
better appearance, texture, and nutrition.
4.3 hIGh-prESSUrE prOCESSING OF MEat aND pOULtrY prODUCtS
Meat is a rich nutrient matrix having high moisture and protein content with neutral pH creating
an ideal environment for growth and proliferation of spoilage microorganisms and food-borne
pathogens. Therefore, adequate processing and preservation technologies have to be employed
in order to preserve its safety, quality, and shelf life. HPP is used as a post-processing step for
improving quality and lengthening shelf life of ready-to-eat meat products (Jofre et al., 2009).
Several studies have been carried out on the effects in HPP of meat and meat products, such as
microbial inactivation (Garriga et al., 2004), texture (Jung et al., 2000), colour changes and struc-
tural changes of myoglobin in minced beef (Carlez et al., 1995) and in pork (Cheah and Ledward,
1997; Wackerbarth et al., 2009), kinetics of the radical formation (Bolumar et al., 2012), and lipid
oxidation in poultry (Kruk et al., 2011) and beef (McArdle et al., 2010). With initial successes in jam
and fruit juice, HPP has growing applications in other food products like smoothies, rice products,
guacamole, salsa, meat, fish, and shellfish (Murchie et al., 2005). Studies on meat, fish, and shrimp
revealed HPP technology as a useful processing technique for muscle foods (Kaur et al., 2013). This
has been used for several meat products of different species origin (Souza et al., 2011). Even though
this technology has many benefits, it encourages the detrimental changes in texture, colour, and
structure and increase in the lipid oxidation levels in meat products (Ma et al., 2007). The changes
caused during HPP of meat and poultry are depicted in Figure 4.2.
Pressures up to 800 MPa and temperatures between 5°C and 40°C are used in commercial appli-
cations (Heinz and Buckow, 2010). Depending on the pressure level and duration, HP processing
of meat may also trigger lipid oxidation (Orlien et al., 2000). It has been recognized that 300 and
600 MPa of pressure levels are critical for encouraging lipid oxidation. The use of antioxidants can
prevent the high pressure-induced lipid oxidation (Mariutti et al., 2008). Most of the vegetative
bacteria can be inactivated by 300–600 MPa pressure (Smelt, 1998). Lopez-Caballero et al. (2000)
Microbes
inactivation
Tissue
disintegration
Lipids
transition
Protein
unfolding
Figure 4.2 Changes during HPP of meat and poultry.
reported that a pressure level of 200–400 MPa at 7°C for 10 min is sufficient for reduction of all tar-
geted microorganisms in prawn. 10–50 MPa pressure levels lower the growth rate of microbes but
higher pressure levels can inactivate the microbes (Rademacher, 2006). Yeasts and moulds are more
sensitive than vegetative bacterial cells to pressure (Patterson, 2005) but ascospores are extremely
resistant to pressure treatments (Chapman et al., 2007). The mechanism for microbial destruction
in HPP is by cell membrane alterations (Moussa et al., 2007), dissociation of ribosome (Abe, 2007),
agglomeration (Farr, 1990), and denaturation of proteins (Barbosa-Canovas et al., 1995).
4.3.1 Impact of hpp on Colour of Meat
Meat colour is controlled by the optical properties of its surface in addition to the myoglobin con-
tent of the muscle. On the other hand, the colour of cured meat products is mainly produced due to the
occurrence of nitrosyl myoglobin, which is produced as a consequence of the reaction of nitric oxide
(from sodium nitrite or sodium nitrate) and myoglobin. HPP is known to aggravate severe modifica-
tions in fresh meat colour although the changes in cured meat products are tolerable and depend on
the moisture content and aw value (Ferrini et al., 2012). HPP treatment increased L* value and reduced
a* and b* values in raw cured hams with high water contents. The HPP treatment had insignificant
effect on the raw cured hams with low water contents. HPP of minced beef increased the L* colour
values significantly in the pressure range 200–350 MPa providing a pink colour to the meat, while
a* values decreased at 400–500 MPa resulting in a grey-brown colour (Carlez et al., 1995). Similar
effects on meat colour have been illustrated by other researchers (Bak et al., 2012a; Reddy et al., 2015).
Generally, high pressure causes remarkable alterations in the colour of fresh meat and thus makes it
difficult for the commercialization of HPP fresh meats as they lack the characteristic colour of fresh
meat from the consumer’s perspective (Cheftel and Culioli, 1997). However, these changes are not
important if the products are further processed into products like sausages, salami, ham, chorizo, etc.
Wackerbarth et al. (2009) employed resonance Raman spectroscopy to show the existence
of a ferrous deoxy-myoglobin after HPP which is undesirable due to the brownish colour, low
before pressurization, a minor conversion to ferric myoglobin was observed in a model system
(Wackerbarth et al., 2009). Pork meat treated above 300 MPa became significantly less red and
more yellow within the first day of storage. This fact was explained by the formation of a short-lived
ferro haemachrome myoglobin species which is transformed into a brown, ferric form of the pig-
ment within the first day of storage (Bak et al., 2012).
Investigations on cured meat products reported an increase in lightness and a decrease in red-
ness when products are pressurized. Colour changes in restructured ham were evaluated and it was
found that the L* and a* values were best preserved in high pH/high salt restructured ham and
had no effect on ham dried to 50% weight loss (Bak et al., 2012). Szerman et al. (2011) studied the
application of HHP on beef carpaccio at three different levels (400, 500, and 600 MPa) with low
temperatures (0°C–5°C) and at room temperature (20°C) and observed that the negative effect on
chromatic parameters was reduced under frozen conditions and also on the water-holding capac-
ity, indicating a minimization of the denaturation of the sarcoplasmic and myofibrillar proteins but
showing a lower inactivation on microorganisms. In another study about the influence of HPP on
marinated poultry products, it was concluded that high pH and addition of sodium carbonate, even
at low concentrations, improved colour and colour stability of products (Schmidgall et al., 2011).
In general, the negative impact of HPP on fresh meat and meat products is dependent on different
parameters, and all of them are not well-understood as of now. In general, HPP colour-induced
changes vary according to the myoglobin content and are more dramatic for fresh red meat than for
white meat and cured meat products. Undesired changes can be limited by optimizing the process
parameters of HPP treatment such as pressure, time, temperature, curing, oxygen removal, and pH.
When looking for a reduction of colour changes induced by HPP, one should keep in mind that mea-
sures to protect the colour quality and stability can result in changed microbial inactivation kinetics
and thus safety and shelf‐life of the final product.
Poultry muscle also undergoes changes in appearance under pressure, although the changes are
less drastic than in mammalian muscle at low pressures (<300 MPa) and temperatures. The pink
colour of fresh turkey breast muscle was altered quickly at pressures higher than 300 MPa at 10°C
(Tintchev et al., 2010). However, it is not as sensitive to HPP as the colour of chicken breast. On the
contrary, Yoshioka et al. (1992) reported that chicken meat only slightly paled after pressurization
at 500 MPa for 10 min.
Increased lightness of meat is a well-documented result of application of HPP on red muscles
(Carlez et al., 1995; Goutefongea et al., 1995; Shigehisa et al., 1991). This whitening effect had been
related either to protein coagulation, which would affect sample structure and surface properties
(Goutefongea et al., 1995), or to globin denaturation and heme group displacement or release (Carlez
et al., 1995). Comparing the pressure levels, meat treated at 600 MPa showed lower a* values than meat
treated at 400 MPa (Carlez et al., 1995). Other authors have observed a reduction of a* values at pres-
sures above 350–400 MPa (Carlez et al., 1995; Jung et al., 2003). The reduction of a* values at higher
pressures has been related to the oxidation of ferrous myoglobin to ferric metmyoglobin and it would
result in the brown colouration of meat observed at those pressures (Carlez et al., 1995). This postulation
would also be consistent with the increase of yellowness (b*) at 400 and 600 MPa. Pressurisation at
200 MPa caused no changes (p > 0.05) in b* values compared to non-treated meat. The temperature of
pressurisation had no significant effect (p > 0.05) on a* and b* values of meat, while higher L* values
were observed in samples pressurised at 30°C than at 10°C (Carlez et al., 1995).
An increase in lightness in raw chevon muscle subjected to HPP of 600 MPa was observed by
Reddy et al. (2015). The appearance of muscle was dependent on pressure treatments making it
whiter with increase in pressures as shown in Figure 4.3. An increase in L* values is the most often
reported modifications occurring in raw meat above the pressure levels of 200 MPa (Del Olmo
et al., 2010; Marcos et al., 2010). The changes in lightness were due to the myofibrillar proteins’
denaturation (Goutefongea et al., 1995) and also myoglobin denaturation (Carlez et al., 1993). It has
been found that an increase in the metmyoglobin content due to pressures leads to decrease in a*
Figure 4.3 Colour changes in raw chevon following high-pressure treatment. (Adapted from reddy, K.J. et al.,
Food Bioprocess Technol., 8, 2347–2358, 2015.)
values (Ledward, 1998). Changes in the colour attributes were also observed during refrigerated
storage for both untreated and treated samples. According to Jung et al. (2003), the changes in
colour of the meat during storage could be associated with both enzymatic and non-enzymatic reac-
tions, resulting in degradation of myofibrillar proteins and disorganization of the myofibrils.
4.3.2 Impact of hpp on Flavour
The effect of HPP on low molecular weight compounds is minimal. Hence vitamins, flavour
compounds, and pigments endure HPP in comparison with conventional processing methods.
Generally, it is assumed that the fresh flavour is retained in pressurised food products, since small
flavour molecules are not affected by high pressures. However, high pressures may affect chemical
and enzymatic reactions resulting in changes in flavour profile. Regarding meat products, major
chemical deteriorative reaction is lipid oxidation, and several investigations have dealt with the
effect of pressure on lipid oxidation in various raw meats, muscles, and meat products (Beltran
et al., 2003, 2004b; Bragagnolo et al., 2007; Campus et al., 2008; Cava et al., 2009; Cheah and
Ledward, 1995, 1996, 1997; Dissing et al., 1997; Ma et al., 2007; Mariutti et al., 2008; Orlien et al.,
2000; Tume et al., 2010; Wiggers et al., 2004). Although different meats and their products show
different effects of high pressure on their oxidative stability, depending on temperature, pressure
level and duration; treatment between 400 and 600 MPa being more critical for oxidative damage
in chicken breast (Ma et al., 2007; Orlien et al., 2000), minced chicken meat (Beltran et al., 2003;
Mariutti et al., 2008), turkey thigh (Dissing et al., 1997), and whole beef muscle (Ma et al., 2007).
It is well known that heat treatment of meat initiates lipid oxidation and enhances development
of off-flavours during long-term storage (Igene and Pearson, 1979; Igene et al., 1985; Kerler and
Grosch, 1996).
During heating, flavour profile of meat is changed and new compounds are produced, primarily
by lipid oxidation and Maillard browning reactions, which together are responsible for the desired
cooked flavour of meat. The changes in the volatile profile of heated meat following pressurisation
(400 MPa) of beef and chicken breast were assessed by Rivas-Cañedo et al. (2009). This study
concluded that pressure treatment affected the levels of some volatile compounds produced by
microbial activity.
Although raw meat has little aroma, it is of significance to explore if HPP preserves the aroma
profile of raw meat, as the released volatile compounds on opening the package should be accept-
able to the consumers and the characteristic flavour attributes have to remain stable for some addi-
tional time during refrigerated storage.
The practical implementation of the high-pressure technology to produce high quality sen-
sorially stable meat products is promising. Lipid oxidation subsequent to off-flavour develop-
ment achieves significance in case of cooked and high pressurised (600 MPa) meat only when
vacuum-packaged meat is re-exposed to oxygen. In contrast to cooked meat, high-pressure-treated

samples showed no (400 MPa) or a considerably delayed (600 MPa) lipid autoxidation when the
meat was re-exposed to oxygen (Schindler et al., 2010). Pressure treatment of beef and chicken
did not induce severe changes of their raw aroma profiles (Schindler et al., 2010). In another study,
the pressurised treatment (400 MPa) reduced the content of volatile compounds, particularly those
derived from Maillard reaction (Campus et al., 2008). HPP treatment increased the overall autolytic
activity of raw meat and leads to a higher concentration of free amino acids (Ohmori et al., 1991).
4.3.3 Impact of hpp on texture
Pressure induces texture modifications by affecting the myofibrillar protein structure and their
gel forming properties. Muscle protein gelation by combined use of high pressure and temperature
was reviewed by Colmenero (2002). The effect of high pressure on the texture of meat and meat
products was reviewed by Sun and Holley (2010). The thermo-labile nature of muscle proteins and
the previously described effect of pressure on the gelation properties of meat and meat proteins will
permit the development of novel meat-based products rooted in an improved structure. Economic
benefits in terms of time and energy savings are associated to the use of HPP as a processing step
for the meat industry.
In fresh meat, the application of low pressure levels has been used to improve the functional
and rheological properties of turkey meat with low pH or PSE meat (Chan et al., 2011). In general,
low pressures (200 MPa) help in tenderizing pre-rigor meat, but tenderization post-rigor with HPP
can only be accomplished by higher temperatures (Sun and Holley, 2010). The influence of HPP
on the meat tenderness depends on the rigor stage, pressure and temperature level applied, and
their combination (Sun and Holley, 2010). Meat tenderization by HPP is likely caused by lysosome
breakdown and subsequent proteolytic activity release to the medium (Hugas et al., 2002). Pre-rigor
treatment of fresh meat by HPP was shown to be very effective to improve the tenderness of fresh
meat. However, the application of HPP at pre-rigor state would require the development of hot bon-
ing at slaughterhouses (Rastogi et al., 2007).
Moreover, the application of HPP can be used to improve the water retention properties of raw
material used for the production of meat products and as a result to the development of products with
reduced salt content (Chan et al., 2011). In this sense, Sikes et al. (2009) made use of high pressure
to reduce the cook loss and to improve the texture of low-salt beef sausage batters. In another study,
HPP was employed for the production of liver sausage, which is a traditional German cooked spread-
able sausage requiring two individual thermal treatments. Due to the high time and temperature
requirements valuable macro- and micro-nutrients are lost (Heinz et al., 2009). These twin thermal
treatments can be replaced by HPP at 600 MPa for 2–5 min at room temperature. The first pressure
treatment of raw material was intended to denature myofibrillar proteins and to produce the desired
product characteristics of consistency and texture, while the second pressure treatment was carried
out after emulsifying the pressurized raw material using raw liver in the bowl chopper to enhance the
shelf-life and to ensure final product characteristics. Patties treated at 200 and 400 MPa exhibited an
improvement in their water holding capacity (8%), cook loss (6%), and textural properties compared
to control (Hygreeva et al., 2016). Other meat products that can benefit from the application or simi-
lar HPP process, with increased quality and improved energy efficiency, are cooked cured products,
such as pork loins or spreadable fermented sausages (Lickert et al., 2010).
4.3.4 Impact of hpp on Lipids
HPP has acknowledged much attention as a novel food preservation method. It is a minimal pro-
cessing technology, which assists as a cold pasteurisation that does not weaken the nutritional and
sensorial characteristics (Rastogi et al., 2007). However, HPP induces lipid oxidation in meat and
meat products. A pressure level of 300–600 MPa is known to be critical for inducing lipid oxidation
(Beltran et al., 2003, 2004a; Mariutti et al., 2008). As a result, the meat develops oxidised flavour which
limits the applicability of HPP in meats. Several studies have revealed that the higher the pressure
and processing time the greater the level of lipid oxidation. Lipid oxidation was significantly acceler-
ated during 24 h of cold storage in both cooked chicken and beef when exposed to oxygen, while the
pressurised and oxygen-exposed chicken and beef meat remained stable (Schindler et al., 2010). The
presence of a threshold around 500 MPa has also been accepted for the lipid oxidation to be induced
(Beltran et al., 2004a; Fuentes et al., 2010). Ma et al. (2007) reported that HPP can lead to alterations
in structure, colour, and oxidation levels of lipids. Bolumar et al. (2011) evaluated the lipid oxidation
of high-pressure-processed (800 MPa, 10 min, 5°C) chicken patties during refrigerated storage (5°C)
and found that lipid oxidation was limited to the surface of the meat patties and active packaging with
10% rosemary extract solution proved effective in inhibiting lipid oxidation and extending its shelf life.
HP may lead to the vulnerability of lipids due to molecular oxygen attack, which can result in the
disruption of fatty acid composition as well (Ma et al., 2007). Studies on the effect of HPP (260, 500,
and 800 MPa for 3–5 min) on the fatty acid profile (PUFA/SFA and n6/n3 ratios) of oysters and salmon
did not show significant changes (Cruz-Romero et al., 2008; Yagiz et al., 2009). Fatty acids play an
important role in several meat quality characteristics and establishing the effect of varied pressure
treatments on fatty acid profile is extremely essential (Wood et al., 2008) in the application of HPP for
meat products.
4.3.5 Impact of hpp on Non-heme Iron
Two mechanisms have been proposed to explain the pressure-induced lipid oxidation: increased
accessibility of iron from heme proteins and membrane disruption. Several studies have observed
that the addition of ethylene diamine tetra acetic acid (EDTA), which can chelate metal ions such as
iron, can be correlated with a reduction of the lipid oxidation in meat processed by HP, which sug-
gests that transition metal ion catalysis is the major factor causing increased lipid oxidation (Beltran
et al., 2004b; Cheah and Ledward, 1997; Ma et al., 2007). However, iron release was not observed
after HP treatment of chicken breast (Orlien et al., 2000). In the same study, it was also concluded
that the catalytic activity of metmyoglobin did not increase during HP-treatment, indicating that
pressure-induced changes of the metmyoglobin conformation that facilitates the access to the cata-
lytic heme group did not take place (Orlien et al., 2000). So far, role of iron in the induction of lipid
oxidation of meats treated by HP is not well-established. Membrane disruption facilitates contact
between unsaturated lipids from the membrane and enzymes and catalysts like heme, non-heme
iron, and other metal cations and, thus, may contribute to the initiation of lipid oxidation. Recently,
the formation of free radicals during HP has been proposed as a possible mechanism behind the
induction of lipid oxidation in HP-processed meats (Bolumar et al., 2011). Radical formation in
the aqueous and lipid phases from HP-treated meat was first reported by Mariutti et al. (2008) and
further studied by Bolumar et al. (2012), who characterised the kinetics of the formation of radicals
in chicken meat during application of different HP treatments. It was found that there is a thresh-
old for the formation of radicals under HP conditions at 400 MPa at 25°C and 500 MPa at 5°C.
However, the chemical mechanism which leads to the formation of radicals in meats by HP is so far
poorly described. Reddy et al. (2015) reported a significant increase (p < 0.05) in non-heme iron in
pressure-treated chevon samples of 300 and 600 MPa. They further concluded that increase in the
non-heme iron with HPP may be due to the opening of the myoglobin porphyrin ring with pressure.
4.3.6 Impact of hpp on proteins
The application of pressure on proteins leads to varied extent of modifications in protein structure.
As a general mechanism, the application of pressure induces unfolding of the protein structure and
subsequent folding after pressure release. This can lead to changes depending on the specific protein
and conditions applied to partial or total denaturation and tuning of electrostatic interactions.
Based on the principle of Le Chatelier and Braun, reactions with a decrease in volume are
favoured by pressure. Denaturation of proteins during HPP is one of the main causes for the inac-
tivation of microbes and microbial inactivation and unalterable changes in muscle proteins start at
a relative level to that necessary for the inactivation of microorganisms. HP treatment affects the
protein conformation of the meat system whereby interesting properties like texture and water-
holding capacity are modified, and thus, it has been recently used as a new dimension in product
development (Sikes et al., 2009; Sun and Holley, 2010).
Covalent bonds have a low compressibility and are much less sensitive to changes in pressure
(Cheftel and Culioli, 1997). HPP induces the breakdown of salt bonds, due to electrostriction and
also parts of hydrophobic interactions. In contrast, hydrogen bonds appear to be slightly strength-
ened under pressure (Cheftel and Culioli, 1997). Quaternary structure is mainly held by hydrophobic
interactions and thus they are very sensitive to pressure (Rastogi et al., 2007). The main alterations
in the tertiary structure are detected beyond 200 MPa and alterations in secondary structure will
take place only at very high pressure above 700 MPa (Rastogi et al., 2007). Muscle proteins includ-
ing myofibrillar proteins are unfolded up to a pressure of 300 MPa. Pressures above this level pro-
duce increased denaturation, gel formation, and agglomeration of proteins. This information can
be in use in product development since improved gel structure and water binding capacity can be
attained by the use of certain HPP treatments (Sun and Holley, 2010).
Modification of the actin-myosin complex is one of the significant effects of high pressure in
meat. Pressure induces structural changes in the main constituent of muscle filaments, which are
possibly caused by increased ATPase activity at 30 MPa as well as with an increase of soluble
materials from the myofibrils enhanced by pressurization above 150 MPa (Nishiwaki et al., 1996).
Suzuki et al. (1990) investigated the effect of high pressure (100–300 MPa) on post-rigor beef mus-
cle for meat tenderization and observed that maximum fragmentation was achieved within 5 min
at 300 MPa. Additionally, the z-line in myofibrils was not apparent in the pressurized muscles. In
contrast, limited effect is visible on connective tissue at ambient temperature and little effect is seen
at higher temperatures (Beilken et al., 1990).
HPP of meat and its products affects protein conformation based on the pressure treatments, hold-
ing time, and temperature, resulting in denaturation and aggregation of proteins (Cheftel and Culioli,
1997). Sarcoplasmic protein fractions are mostly labile proteins of post-mortem muscle (Lawrie,
1998). A mix of several hundred globular proteins of comparatively low molecular weight is identified
to be present in the sarcoplasmic fraction (Tornberg, 2005). Sarcoplasmic protein denaturation proved
to deliver an impact on meat quality attributes like water holding capability and colour (Sayd et al.,
2006). Moreover, these proteins have a role in the consistency of cooked meat (Farouk et al., 2002).
4.3.7 Impact of hpp on thermal Behaviour
Differential scanning calorimetry (DSC) is a powerful technique that has been used to study the
structural and thermal properties of natural polymers such as proteins and it has been used to relate
the denaturation of individual muscle proteins to the textural changes in meat caused by cooking
(Findlay et al., 1986) and pressurization (Angsupanich and Ledward, 1998; Angsupanich et al.,
1999). The three major endothermic transitions seen in beef muscle, attributed to myosin, collagen,
and actin, have been associated with specific changes in meat texture (Findlay et al., 1986). Effect
of HPP in chicken nuggets was evaluated by Feby et al. (2015) and the DSC thermograms of control
and pressurized samples obtained from the studies are depicted in Figure 4.4. Cooked chicken meat
showed transitions at 60°C (myosin), 71°C (sarcoplasmic and connective tissue proteins), and 81°C
(actin) with melting enthalpy of 1.16 kJ·kg−1, 2.8 kJ·kg−1, and 2.5 kJ·kg−1, respectively. The typical
transition temperature for myosin is 59°C, for sarcoplasmic proteins is 69°C, and for actin is 80°C
Figure 4.4 Differential scanning calorimetry profiles of low-sodium re-structured chicken nuggets subjected to
high-pressure treatment. (Adapted from Feby et al., J Food and Nutr Res., 54(4): 334–345, 2015.)
as observed by other researchers in chicken muscle (Amako and Xiong, 2001; Jiménez-Colmenero
et al., 1998; Kijowski and Mast, 1988; Trespalacios and Pla, 2007a, 2007b). In control nuggets,
peaks at 55°C, 62°C, and 70°C with an associated melting enthalpy of 1.2 kJ·kg−1, 0.7 kJ·kg−1, and
2.1 kJ·kg−1, respectively, which tentatively could be attributed to the thermal denaturation of native
proteins myosin (first transition) and actin (last transitions) of residual actomyosin complex, left over
after heating. The intermediate transitions are mainly due to sarcoplasmic and connective proteins.
Several fused peaks around 70°C–90°C were recorded with a maximum transition temperature
of 85.93°C and ∆H of 1.02 kJ·kg−1. The shift in myosin and actin peaks observed in the nuggets
could be due to destabilizing effects of sodium chloride and potassium chloride on myofibrillar
proteins (Kijowski and Mast, 1988). It was reported that actin shows high sensitivity (reduction
in melting temperature up to 16°C) in the presence of salts like NaCl (Barbut and Findlay, 1991).
Moreover, interaction between whey protein and myofibrillar proteins during heating can cause a
downward shift in myosin and actin endothermic peaks and, in some cases, complete disappearance
of the actin peak (Liu et al., 2000). With increasing pressure levels, proteins became denatured and
endothermal peaks decreased in size or were absent in the thermogram. In the case of 200 MPa-
processed sample, these peaks were seen at 53.39°C, 60.95°C, and 69.8°C but with reduced enthal-
pies of denaturation (0.8, 0.6, and 1.25 kJ·kg−1, respectively). Only sarcoplasmic and actin peaks
(above 70°C) were seen in 400 MPa-processed sample (with melting enthalpy of 0.98 kJ·kg−1), while
the endothermic peaks disappeared in 600 MPa-processed nuggets. Actin is generally considered
to be more stable to heating and pressure treatments. Similar results were earlier reported in studies
on pressure treatment of pork and duck meat batters after heating (Fernández-Martin et al., 1997;
Khan et al., 2014). It can be noted that endothermic events centred around 80°C–90°C seen in all
samples, which can be assigned to β-lactoglobulin and α-lactalbumin, the two major whey protein
components (Liu et al., 2000), survived pressure treatment. This was confirmed by the DSC trace of
whey protein concentrate (4.1% moisture) (Feby et al., 2015) which showed a maximum transition
temperature at 87.99°C ΔH = 151 kJ·kg−1.
4.3.8 Impact of hpp on Water holding Capacity
Water holding capacity plays an important role as a physical quality marker both in fresh and
processed meat products. The market value and consumer acceptability are primarily influenced
due to this parameter. It has direct impact on palatability characteristics such as texture, appearance,
juiciness, and overall eating quality (Hughes et al., 2014).
There is a lack of clarity in the literature on the influence of high pressure on water-holding char-
acteristics of meat and poultry products, and the literature is contradictory. Positive, negative, and no
effect reports on the effects of HPP on water-holding capacity are reported which may be due to the
influential effect of varied products nature, processing pressure-holding, time-temperature, and the
ingredients used. Intelligent combination of pressure, temperature, and proper natural meat additives
can substantially improve the WHC in meat and poultry products during HPP (Yang et al., 2015). The
pressurized conditions may unfold meat proteins especially myosin, which lead to the formation of
flexible structures which can hold more moisture content. Chattong and Apichartsrangkoon (2009)
evaluated the effect on WHC of meat at varied pressure levels, time, and temperature and reported
the improvement in WHC at higher severity levels of processing. So, further research is required to
understand the mechanism of the effect of HPP on the WHC of meat and poultry products and design-
ing of proper combination processing techniques with natural additives may solve the problem.
4.4 EFFECt OF hpp ON MICrOOrGaNISMS
As a food preservation method, the effectiveness of HPP in destroying food-borne microorgan-

isms depends on intrinsic and extrinsic factors. HPP of foods results in changes in the cell mem-
branes, cell wall, proteins, and enzyme-mediated cellular functions which lead to the inactivation of
microorganisms (Simpson and Gilmour, 1997). The response of vegetative pathogenic and spoilage
microorganisms in meat and meat products to HPP is variable and depends on process parameters
such as pressure, temperature, and processing time and on product parameters such as pH, aw, salt
content, and the presence of other antimicrobials (Rendueles et al., 2011; Toepfl and Heinz, 2009).
The basic principles of HPP on microbial inactivation are based on protein denaturation (Barbosa-
Canovas et al., 1995) and agglomeration of cellular proteins (Farr, 1990). Generally, it is anticipated
that gram-negatives and cells in the growth phase are more sensitive than gram positives and cells
in the stationary phase, respectively. Pressure levels of about 10–50 MPa can decrease the growth
rate and reproduction, whereas, higher pressure levels lead to inactivation (Rademacher, 2006). The
impact of different pressure levels on microorganisms is illustrated in Figure 4.5. The effect of pro-
cess modifications with respect to pressure, temperature, and holding time on different target organ-
isms in HPP of meat products has been depicted in Table 4.1, which illustrates the initial counts
and the reduction achieved through the application of variation in HPP with respect to various meat
products. The data given in Table 4.1 clearly substantiate the literature available on inactivation of
microorganisms in different HPP meat products.
Figure 4.5 effect of HPP on micro-organisms.

table 4.1 Effect of hpp on Inactivation of Microflora in Meat products
hpp processing Microbial reduction

Meat product Conditions (log CFU/g) references
raw meat and Marinated beef 600 MPa at inoculated LAb: about Jofre et al. (2009)
products with loin 31°C for 6 min 4 of 5 initially present
higher aw raw beef 560 MPa at Aerobic mesophiles: Jung et al. (2003)
10°C for 4 min 2.5 the day after
processing
Mechanically 450 MPa at Aerobic mesophiles: yuste et al. (2001)
recovered poultry 20°C for 15 min 3.7 the day after
processing
Dry-cured Dry-cured ham 600 MPa at inoculated LAb: 1.6 of Jofre et al. (2009)
products 31°C for 6 min 4 initially present
Dry-cured chorizo 350 MPa at Aerobic mesophiles: ruiz-Capillas
sausages 20°C for 15 min <1 immediately after et al. (2007a)
processing
Dry-cured beef 500 MPa at Aerobic mesophiles: rubio et al. (2007)
18°C for 5 min 1.66 after processing
2.55 after 60 d of storage
Cooked meat sliced cooked 400 MPa at inoculated Listeria Aymerich et al.
and products ham 17°C for 10 min monocytogenes: (2005)
(low acid, high 1.8 immediately after
aw) processing
blood sausages 600 MPa at Aerobic mesophiles: Diez et al. (2008)
15°C for 10 min 2.62 the day after
processing
Frankfurters 400 MPa at total viable count: ruiz-Capillas
30°C for 10 min 2.16 immediately after et al. (2007b)
processing
4.5 appLICatIONS OF hpp IN MEat, pOULtrY, aND FISh
The most widely used commercial applications for HPP are chilled entrees, delicatessen meats,
and marinated meats, with all but the last in-package processing. HPP can also be used for non-
thermal processing of cured ham. It is employed in the meat and poultry industry to pasteurize
sliced ham, turkey, chicken, and chicken sausages. It can be used for pre-packed meat products to
eliminate the handling and other microbiological contaminations arising during the process. Other
applications include meat tenderization, infusing meats with marinades, and using HPP with bulk
packaging to curb contamination between processing plants. HPP is also exploited in improving
the rheological and functional properties (Alba et al., 2012; Tornberg et al., 1997). HPP is used
to improve the digestibility of meat proteins (Kaur et al., 2016). The apparent biological value of
protein and protein efficiency ratio (PER) of pressure-treated meat are equivalent to meat proteins
tenderized at atmospheric pressure. It also causes inactivation of enzymes while retaining nutrients
and flavour. The fermentation reactions are known to be retarded at high pressures.
Meat and seafood safety and quality can be considerably enhanced by the application of this
technology by eradicating the food borne pathogens (particularly Listeria monocytogenes) in the
post-packaging step. HPP has been effectively demonstrated for enhancing the storage period
of tuna at low temperature (Kamalakanth et al., 2011). Application of HPP results in protein
denaturation, which can inhibit some inherent enzymatic activities and biogenic activity of some
microorganisms in fish and fish products. However, HPP has been shown to accelerate lipid oxi-
dation in fish muscles. Recent intensive research on the effects of high hydrostatic pressure on
fish tissues has gradually revealed the benefits and defects of this novel processing technology
(Ohshima et al., 1993). It has been used in preserving the quality and inactivation of pathogens
in many seafood items, such as oysters, mullet, salmon, cod, squid, and shrimp (Erkan et al.,
2010; Guo et al., 2010; Kaur et al., 2013; Lakshmanan et al., 2003; Ma and Su, 2011; Montiel
et al., 2012). The applicability of this process in seafood industry is gaining importance due to
its ability to inactivate bacterial pathogens and viruses in shellfish and to increase the extraction
yield. The processing efficiency and product yield w.r.t. seafood, especially crustaceans, can be
improved by applying HPP to shuck lobsters and crabs completely from the shell and can create
new markets (Campus, 2010).
4.6 COMBINED trEatMENtS WIth hIGh-prESSUrE prOCESSING
Application of combined hurdles together with HPP has been proposed to increase the micro-
bicidal effect of low-pressure processes in order to minimize the unwanted changes induced by
ultra-high hydrostatic pressures (above 400 MPa) in meat, poultry, and fish products. The undesir-
able effects of HPP could be eliminated by the application of combined treatments. Several treat-
ments are being combined with high pressure; these include low temperature, CO2, ultrasound,
gamma irradiation, E-beam irradiation, natural antimicrobials, natural antioxidants, new packaging
systems, etc. (Garriga and Aymerich, 2009; Hygreeva and Pandey, 2016). Several examples of com-
binations of HPP treatments with additional preservation methods are listed in Table 4.2. Moreover,
additional hurdles or processes are useful to avoid the recovery of sub-lethal injured cells (Jofré
et al., 2010; Liu et al., 2012).
table 4.2 Combinations of hpp treatments with additional preservation Methods

products processing Conditions Effects references
Chicken 300, 600, and 800 MPa treatment at 300 MPa did not induce Alves et al. (2012)
breast meat significant lipid oxidation during
storage period, while 600 and
800 MPa enhanced formation of
secondary lipid oxidation products.
Chicken 300 MPa (5 min, 20°C), synergistic action of HPP with rodríguez Calleja
breast fillet liquid antimicrobial antimicrobial coating in MAP extended et al. (2012)
edible coating, MAP the durability of chicken breast fillets,
packaging which maintained their sensory and
microbiological qualities for up to
28 days.
Chicken 300, 450, and 600 MPa Pressures of 450 and 600 MPa Kruk et al. (2011)
breast fillet (5 min, 15°C) almost completely eliminated
Salmonella, Escherichia, and Listeria.
increasing pressure increased cooking
loss and colour by increasing lightness
(L*), redness (a*), and yellowness (b*)
values and pressure of 450 MPa and
higher induced lipid oxidation.
(Continued)
table 4.2 (Continued) Combinations of hpp treatments with additional preservation Methods
beef and 0.1–800 MPa Pressure treatment of beef samples at Ma et al. (2007)
chicken room temperature led to increase in
muscle tbArs number after 7 days storage at
4°C and this increase was more
noticeable after treatment at pressures
≥400 MPa than after treatment at lower
pressures. similar results were found in
those treated at 40°C, but at 60°C and
70°C, pressure had little additional effect
on the oxidative stability of the muscle.
Pressure treatments of 600 and
800 MPa, at all temperatures, induced
increased rates of lipid oxidation in
chicken muscle, but in general chicken
muscle was more stable than beef to
pressure.
beef 50–600 MPa Pressure higher than 300 MPa induces Jung et al. (2003)
(20–300 s, 10°C) modifications in meat colour
parameters, a decrease in total
microflora, and 1-week delay before
microbial growth (520 MPa, 260 s).
redness of the 520MPa-treated samples
decreases gradually in relation to the
increase of metmyoglobin.
beef sausage 400 MPa HPP produced significant improvement sikes et al. (2009)
batter (2 min at 10°C) in the moisture retention of the cooked
products.
Hardness and gumminess of pressure-
treated samples was higher compared
to untreated samples.
slight increase in “whiteness” with
pressure treatment.
Greater acceptability in appearance and
texture was noted in pressure-treated
sausages of lower salt content
compared with non-pressure-treated
samples.
Cooked ham 400 MPa (10 min at HPP resulted in the reduction of 3–4 log Marcos et al. (2008)
17°C), antimicrobial units of L. monocytogenes
alginate films Combination of antimicrobial packaging
and HPP extended storage life up to
60 days.
“Chorizo” 350 MPa (15 min) HP treatment reduced the level of lactic ruiz-Capillas et al.
(spanish acid bacteria by <1 log CFu/g and (2007a)
dry-cured production of biogenic amines.
sausage)
Low-acid 300 MPa (10 min, 17°C), Application of HPP as an additional Marcos et al. (2005)
fermented three-strain cocktail of hurdle to the ripening process produced
sausage (fuet Salmonella and L. a greater decrease in the Salmonella
and chorizo) monocytogenes population, showing lower counts
(3 MPn/g) in ripened sausages.
A discolouration of sausages was
observed, coinciding with an increase
in L* values of raw sausages
pressurized at 300 MPa.
(Continued)
table 4.2 (Continued) Combinations of hpp treatments with additional preservation Methods
Cooked 500 MPa (5 or 15 min, Pressurized sausages were more Mor-Mur and
sausages 65°C), conventional cohesive and less firm than heat-treated yuste (2003)
heat pasteurisation sausages.
(80°C–85°C for Pressure heat treatment induced higher
40 min), vacuum yield than heat treatments.
packaging Pressurized samples were preferred by
sensory panellists because of their
better appearance, taste, and texture.
beef 60°C and 300 and increase in tenderness rusman et al. (2007)
400 MPa
Frankfurter 60°C and 600 MPa improved gelation tintchev et al. (2013)
batters
Chicken and 200 MPa for 30 min Limited the gelling process and lowered Jiménez Colmenero
pork batters at 70°C firmness et al. (1998)
Dry-cured ham 600 MPa for 6 min equivalent to untreated products Hugas et al. (2002)
at 31°C
rte-cured HHP & bio preservatives Control of L. monocytogenes during Hereu et al. (2012)
meat products storage
Chicken breast HPP & gamma inactivation of C. sporogenes Crawford et al. (1996)
irradiation
Frankfurters 150 and 300 MPa salt HPP at 150 MPa decreased cook loss Crehan et al. (2000)
levels (1.5% and 2.5%) values and increased stability of meat
emulsions at low salt levels.
texture profile analysis values were
also improved after pressure treatment.
Marinated e-beam irradiation shelf life extension of marinated Alahakoon et al. (2015)
chicken breast (1 & 2kGy) & HPP chicken breast
(300 & 400 MPa)
Mutton patties HPP & gamma shelf life extension banerjee et al. (2017)
irradiation
Dry-cured ham HP Co2 50°C, 12 MPa inactivation of C. sporogenes Ferrentino et al.
for 15 min (2012)
4.7 SUMMarY
Out of the novel non-thermal preservation technologies, HPP has been forecasted as a conve-
nient preservation technique for high-quality chemical-free meat, poultry, and fish products. This
technology provides fresh products with natural flavours and taste as it is demanded by consumers.
The effective standardization of pressures can result to benefit microbial safety with minimal
effects on nutritional and sensory attributes. This is a useful post-packaged preservation technology
for ready-to-eat whole or sliced meat, poultry, and fish products. Microbial safety is achieved by
the combination of breakdown of non-covalent bonds and the puncturing or permeabilization of the
cell membrane. The application of HPP has been studied in several meat, poultry, and fish products
and the varied effects of pressure, temperature, and holding time have been determined. HPP has
several advantages and few drawbacks when we think about application and commercialization in
the meat industry. It offers the opportunities of maintaining the fresh-like quality and has got wide
applications in acid foods as well as the retention of macro and micro nutrients. Even though excel-
lent microbial safety can be achieved by standardizing the processing parameters, lipid oxidation
at higher pressures especially in meat, poultry, and fish products is a problem to be addressed.
This can be curtailed by incorporation of suitable antioxidants or by other combination techniques.
Standardization of proper protocols in terms of combination techniques and natural additives may
definitely result in commercialization of this technology, which will give beneficial results in the
meat and seafood industry.
rEFErENCES
Abe, F. (2007). Exploration of the effects of high hydrostatic pressure on microbial growth, physiology and
survival: Perspectives from piezophysiology. Biosci Biotechnol Biochem. 71: 2347–2357.
Alahakoon, A.U., Jayasena, D.D., Jung, S., Kim, S.H., Kim, H.J., and Jo, C. (2015). Effect of electron beam
irradiation and high pressure treatment combined with citrus peel extract on seasoned chicken breast
meat. J. Food Process Pres. 39: 2332–2339.
Alba, M.D., Montiel, R., Bravo, D., Gaya, P., and Medina, M. (2012). High pressure treatments on the inactiva-
tion of Salmonella enteritidis and the physico-chemical, rheological and colour characteristics of sliced
vacuum-packaged dry-cured ham. Meat Sci. 91: 173–178.
Alves, A.B., Bragagnolo, N., da Silva, M.G., Skibsted, L.H., and Orlien, V. (2012). Antioxidant protection of
high-pressure processed minced chicken meat by industrial tomato products. Food Bioprod Process. 90:
499–505.
Amako, D.E., and Xiong, Y.L. (2001). Effects of carrageenan on thermal stability of proteins from chicken
thigh and breast muscles. Food Res Int. 34: 247–253.
Angsupanich, K., and Ledward, D. A. (1998). High pressure treatment effects on cod (Gadus morhua) muscle.
Food Chem. 63(1): 39–50.
Angsupanich, K., Edde, M., and Ledward, D. A. (1999). Effects of high pressure on the myofibrillar proteins
of cod and Turkey muscle. J Agric Food Chem. 47(1): 92–99.
Aymerich, T., Jofré, A., Garriga, M., and Hugas, M. (2005). Inhibition of Listeria monocytogenes and Salmonella
by natural antimicrobials and high hydrostatic pressure in sliced cooked ham. J. Food Prot. 68: 173–177.
Bak, K.H., Lindahl, G., Karlsson, A.H., and Orlien, V. (2012a). Effect of high pressure, temperature, and stor-
age on the colour of porcine longissimus dorsi. Meat Sci. 92: 374–381.
Bak, K.H., Lindahl, G., Karlsson, A.H., Lloret, E., Ferrini, G., Arnau, J., and Orlien, V. (2012b). High pressure
effect on the colour of minced cured restructured ham at different levels of drying, pH, and NaCl. Meat
Sci. 90: 690–696.
Banerjee, R., Jayathilakan, K., Chauhan, O.P., Naveena, B.M., Devatkal, S., and Kulkarni, V.V. (2017).
Vacuum packaged mutton patties: Comparative effects of high pressure processing and irradiation.
J Food Process Pres. 41: 1–9.
Barbosa-Canovas, G.V., Pothakamury, U.R., and Swanson, B.G. (1995). State of the art technologies for the
stabilization of foods by non-thermal processes: Physical methods. In: Food Preservation by Moisture
Control, Barbosa-Cánovas, G.V., and Welti-Chanes, J., Eds. Tecnomic Publishing Co, Lancaster, PA.
Barbut, S., and Findlay, C.J. (1991). Influence of sodium, potassium and magnesium chloride on thermal prop-
erties of beef muscle. J Food Sci. 56: 180–182.
Beilken, S.L., Macfarlane, J.J., and Jones, P.N. (1990). Effect of high pressure during heat treatment on the
Warner–Bratzler shear force values of selected beef muscles. J Food Sci. 55: 15–18.
Beltran, E., Pla, R., Capellas, M., Yuste, J., and Mor‐Mur, M. (2004a). Lipid oxidation and colour in pressure‐
and heat‐treated minced chicken thighs. J Sci Food Agric. 84: 1285–1289.
Beltran, E., Pla, R., Yuste, J., and Mor-Mur, M. (2004b). Use of antioxidants to minimize rancidity in pressur-
ized and cooked chicken slurries. Meat Sci. 66: 719–725.
Beltran, E., Pla, R., Yuste, J., and Mor-Mur, M. (2003). Lipid oxidation of pressurized and cooked chicken:
Role of sodium chloride and mechanical processing on TBARS and hexanal values. Meat Sci. 64: 19–25.
Bolumar, T., Andersen, M.L., and Orlien, V. (2011). Antioxidant active packaging for chicken meat processed
by high pressure treatment. Food Chem. 129: 1406–1412.
Bolumar, T., Skibsted, L.H., and Orlien, V. (2012). Kinetics of the formation of radicals in meat during high
pressure processing. Food Chem. 134: 2114–2120.
Bragagnolo, N., Danielsen, B., and Skibsted, L.H. (2007). Rosemary as antioxidant in pressure processed
chicken during subsequent cooking as evaluated by electron spin resonance spectroscopy. Innov Food
Sci and Emerg Technol. 8: 24–29.
Campus, M. (2010). High pressure processing of meat, meat products and seafood. Food Eng. Rev. 2: 256–273.
Campus, M., Flores, M., Martínez, A., and Toldrá, F. (2008). Effect of high pressure treatment on colour,
microbial and chemical characteristics of dry cured loin. Meat Sci. 80: 1174–1181.
Carlez, A., Rosec, J.-P., Richard, N., and Cheftel, J.-C. (1993). High pressure inactivation of Citrobacter freun-
dii, Pseudomonas fluorescens and Listeria innocua in inoculated minced beef muscle. LWT-Food Sci
Technol. 26: 357–363.
Carlez, A., Veciana-Nogues, T., and Cheftel, J.C. (1995). Changes in colour and myoglobin of minced beef
meat due to high pressure processing. LWT-Food Sci Technol. 28: 528–538.
Cava, R., Ladero, L., Gonzalez, S., Carrasco, A., and Ramirez, M.R. (2009). Effect of pressure and holding
time on colour, protein and lipid oxidation of sliced dry-cured Iberian ham and loin during refrigerated
storage. Innov Food Sci Emerg Technol. 10: 76–81.
Chan, J.T.Y., Omana, D.A., and Betti, M. (2011). Application of high pressure processing to improve the func-
tional properties of pale, soft, and exudative (PSE)-like turkey meat. Innov Food Sci Emerg Technol.
12: 216–225.
Chapman, B., Winley, E., Fong, A.S.W., Hocking, A.D., Stewart, C.M., and Buckle, K.A. (2007). Ascospore
inactivation and germination by high pressure processing is affected by ascospore age. Innov Food Sci
Chattong, U., and Apichartsrangkoon, A. (2009). Dynamic viscoelastic characterisation of ostrich-meat yor
(Thai sausage) following pressure, temperature and holding time regimes. Meat Sci. 81: 426–432.
Cheah, P.B., and Ledward, D.A. (1995). High-pressure effects on lipid oxidation. J Am Oil Chem Soc. 72:
1059–1063.
Cheah, P.B., and Ledward, D.A. (1996). High pressure effects on lipid oxidation in minced pork. Meat Sci.
43: 123–134.
Cheah, P.B., and Ledward, D.A. (1997). Inhibition of metmyoglobin formation in fresh beef by pressure treat-
ment. Meat Sci. 45: 411–418.
Cheftel, J.C., and Culioli, J. (1997). Effects of high pressure on meat: A review. Meat Sci. 46: 211–236.
Colmenero, F.J. (2002). Muscle protein gelation by combined use of high pressure/temperature. Trends Food
Sci Technol. 13: 22–30.
Crawford, Y.J., Murano, E.A., Olson, D.G., and Shenoy, K. (1996). Use of high hydrostatic pressure and irra-
diation to eliminate Clostridium sporogenes spores in chicken breast. J Food Prot. 59: 711–715.
Crehan, C.M., Troy, D.J., and Buckley, D.J. (2000). Effects of salt level and high hydrostatic pressure process-
ing on frankfurters formulated with 1.5 and 2.5% salt. Meat Sci. 55: 123–130.
Cruz-Romero, M., Kerry, J.P., and Kelly, A.L. (2008). Changes in the microbiological and physicochemical qual-
ity of high-pressure-treated oysters (Crassostrea gigas) during chilled storage. Food Cont. 19: 1139–1147.
Del Olmo, A., Morales, P., A’vila, M., Calzada, J., and Nunez, M. (2010). Effect of single-cycle and multiple-
cycle high-pressure treatments on the colour and texture of chicken breast fillets. Innov Food Sci Emerg
Technol. 11: 441–444.
Diez, A.M., Urso, R., Rantsiou, K., Jaime, I., Rovira, J., and Cocolin, L. (2008). Spoilage of blood sausages
Morcilla de Burgos treated with high hydrostatic pressure. Intl J Food Microbiol. 123: 246–253.
Dissing, J., Bruun-Jensen, L., and Skibsted, L.H. (1997). Effect of high pressure treatment on lipid oxidation
in turkey thigh muscle during chill storage. Eur Food Res Technol. 205: 11–13.
Erkan, N., Üretener, G., and Alpas, H. (2010). Effect of high pressure (HP) on the quality and shelf life of red
mullet (Mullus surmelutus). Innov Food Sci Emerg Technol. 11: 259–264.
Farouk, M.M., Wieliczko, K., Lim, R., Turnwald, S., and MacDonald, G.A. (2002). Cooked sausage batter
cohesiveness as affected by sarcoplasmic proteins. Meat Sci. 61: 85–90.
Farr, D. (1990). High pressure technology in the food industry. Trends Food Sci Technol. 1: 14–16.
Feby L., Pandey, M.C., Chauhan, O.P., Khudsia S., and Abhishek, V. (2015). Effect of high pressure processing
on the quality characteristics and shelf life of low-sodium re-structured chicken nuggets. J Food and
Nutr Res. 54(4): 334–345.
Fernández-Martín, F., Fernández, P., Carballo, J., and Jiménez-Colmenero, F. (1997). Pressure/heat combina-
tions on pork meat batters: Protein thermal behavior and product rheological properties. J Agric Food
Chem. 45: 4440–4445.
Ferrentino, G., Balzan, S., and Spilimbergo, S. (2012). Supercritical carbon dioxide processing of dry cured
ham spiked with Listeria monocytogenes: Inactivation kinetics, colour, and sensory evaluation. Food
Ferrini, G., Comaposada, J., Arnau, J., and Gou, P. (2012). Colour modification in a cured meat model dried by
Quick-Dry-Slice process® and high pressure processed as a function of NaCl, KCl, K-lactate and water
contents. Innov Food Sci Emerg Technol. 13: 69–74.
Findlay, C.J., Parkin, K.L., and Stanley, D.W. (1986). Differential scanning calorimetry can determine kinetics
of thermal denaturation of beef muscle proteins. J Food Biochem. 10: 1–15.
Fuentes, V., Ventanas, J., Morcuende, D., Estévez, M., and Ventanas, S. (2010). Lipid and protein oxidation
and sensory properties of vacuum-packaged dry-cured ham subjected to high hydrostatic pressure. Meat
Sci. 85: 506–514.
Garriga, M., and Aymerich, T. (2009). Advanced decontamination technologies: High hydrostatic pressure on
meat products. In: Safety of Meat and Processed Meat, pp. 183–208. Toldrá, F. Ed., Springer, New York.
Garriga, M., Grebol, N., Aymerich, M.T., Monfort, J.M., and Hugas, M. (2004). Microbial inactivation after
high-pressure processing at 600 MPa in commercial meat products over its shelf life. Innov Food Sci
Goutefongea, R., Rampon, V., Nicolas, N., and Dumont, J.P. (1995). Meat colour changes under high pressure
treatment. In: Association, A.M.S. (Ed.), Proceedings of the 41st ICoMST. American Meat Science
Association, p. 384–385.
Guo, J., Lee, H.Y., and Ahn, J. (2010). Effect of high pressure processing on the quality of squid (Todarodes
pacificius) during refrigerated storage. Food Chem. 119: 471–476.
Heinz, V., and Buckow, R. (2010). Food preservation by high pressure. J Verbrauch Lebensm, 5: 73–81.
Heinz, V., Knoch, A., and Lickert, T. (2009). Product innovation by high pressure processing. New Food. 2: 43–47.
Hereu, A., Bover-Cid, S., Garriga, M., and Aymerich, T. (2012). High hydrostatic pressure and biopreservation
of dry-cured ham to meet the food safety objectives for Listeria monocytogenes. Int J Food Microbiol.
154: 107–112.
Hughes, J.M., Oiseth, S.K., Purslow, P.P., and Warner, R.D (2014). A structural approach to understanding the
interactions between colour, water holding capacity and tenderness. Meat Sci. 98: 520–532.
Hygreeva, D., and Pandey, M.C. (2016). Novel approaches in improving the quality and safety aspects of processed
meat products through high pressure processing technology: A review. Trends Food Sci Technol. 54: 1–11.
Hygreeva, D., Pandey, M.C., and Chauhan, O.P. (2016). Effect of high-pressure processing on quality char-
acteristics of precooked chicken patties containing wheat germ oil wheat bran and grape seed extract.
J Food Process Pres. doi:10.1111/jfpp.12980.
Igene, J.O., and Pearson, A.M. (1979). Role of phospholipids and triglycerides in warmed-over flavor develop-
ment in meat model systems. J Food Sci. 44: 1285–1290.
Igene, J.O., Yamaguchi, K., Pearson, A.M., and Gray, J.I. (1985). Mechanism by which nitrite inhibits the
development of warmed over flavour in cured meat. Food Chem. 18: 1–18.
Jiménez-Colmenero, F., and Borderias, A.J. (2003). High-pressure processing of myosystems. Uncertainties
in methodology and their consequences for evaluation of results. Eur Food Res Technol. 217: 461–465.
Jiménez-Colmenero, F., Cofrades, S., Carballo, J., Fernández, P., and Fernández-Martín, F. (1998). Heating
of chicken and pork meat batters under pressure conditions: Protein interactions. J Agric Food Chem.
46: 4706–4711.
Jofré, A., Aymerich, T., Bover-Cid, S., and Garriga, M. (2010). Inactivation and recovery of Listeria monocy-
togenes, Salmonella enterica and Staphylococcus aureus after high hydrostatic pressure treatments up
to 900 MPa. Int Microbiol. 13: 497–503.
Jofre, A., Aymerich, T., Grèbol, N., and Garriga, M. (2009). Efficiency of high hydrostatic pressure at 600 MPa
against food-borne microorganisms by challenge tests on convenience meat products. LWT-Food Sci
Technol. 42: 924–928.
Jung, S., Ghoul, M., and De Lamballerie-Anton, M. (2000). Changes in lysosomal enzyme activities and shear
values of high pressure treated meat during ageing. Meat Sci. 56: 239–246.
Jung, S., Ghoul, M., and De Lamballerie-Anton, M. (2003). Influence of high pressure on the colour and
microbial quality of beef meat. LWT-Food Sci Technol. 36: 625–631.
Kamalakanth, C.K., Ginson, J., Bindu, J., Venateswarlu, R., Das, S., Chauhan, O.P., and Gopal, T.K.S. (2011).
Effect of high pressure on K-value, microbial and sensory characteristics of yellowfin tuna (Thunnus
albacares) in EVOH films during chill storage. Innov Food Sci Technol. 12: 451–455.
Kaur, L., Astruc, T., Vénien, A., Loison, O., Cui, J., Irastorza, M., and Boland, M. (2016). High pressure pro-
cessing of meat: Effects on ultrastructure and protein digestibility. Food Funct. 18: 2389–2397.
Kaur, B.P., Kaushik, N., Rao, P.S., and Chauhan, O.P. (2013). Effect of high-pressure processing on physi-
cal, biochemical, and microbiological characteristics of black tiger shrimp (Penaeus monodon). Food
Kerler, J., and Grosch, W. (1996). Odorants contributing to warmed-over-flavour (WOF) of refrigerated cooked
beef. J Food Sci. 61: 1271–1275.
Khan, M.A., Ali, S., Abid, M., Cao, J., Jabbar, S., Tume, R.K., and Zhou, G. (2014). Improved duck meat
quality by application of high pressure and heat: A study of water mobility and compartmentalization,
protein denaturation and textural properties. Food Res Int. 62: 926–933.
Kijowski, J.M., and Mast, M.G. (1988). Effect of sodium chloride and phosphates on the thermal properties of
chicken meat proteins. J Food Sci. 53: 367–370.
Kruk, Z.A., Yun, H., Rutley, D.L., Lee, E.J., Kim, Y.J., and Jo, C. (2011). The effect of high pressure on micro-
bial population, meat quality and sensory characteristics of chicken breast fillet. Food Cont. 22: 6–12.
Lakshmanan, R., Piggott, J.R., and Paterson, A. (2003). Potential applications of high pressure for improve-
ment in salmon quality. Trends Food Sci Technol. 14: 354–363.
Lawrie, R.A. (1998). The conversion of muscle to meat. In: Lawrie’s Meat Science. 6th ed. pp. 96–118.
Woodhead Publishing, Cambridge, UK.
Ledward, D.A. (1998). High pressure processing of meat and fish. In: Fresh Novel Foods by High Pressure,
pp. 165–175. Autio K. Ed., VTT Biotechnology and Food Research, Espoo, Finland.
Lickert, T., Badewien, M., Vorwold, G., Töpfl, S., Knoch, A., and Heinz, V. (2010). Herstellung neuartig inno-
vativer Fleisch- und Wurstprodukte mittels hoher hydrostatischer Drücke—The production of novel
innovative meat products by high hydrostatic pressure. Fleisch. 6: 56–59.
Liu, Y., Betti, M., and Gänzle, M.G. (2012). High pressure inactivation of Escherichia coli,Campylobacter
jejuni, and spoilage microbiota on poultry meat. J Food Prot. 75: 497–503.
Liu, G., Xiong, Y.L., and Butterfield, D.A. (2000). Chemical, physical and gel-forming properties of oxidized
myofibrils and whey- and soy-protein isolates. J Food Sci. 65: 811–818.
Lopez-Caballero, M.E., Pérez-Mateos, M., Borderias, J.A., and Montero, P. (2000). Extension of the shelf life of
prawns (Penaeus japonicus) by vacuum packaging and high-pressure treatment. J Food Prot. 63: 1381–1388.
Ma, H.J., Ledward, D.A., Zamri, A.I., Frazier, R.A., and Zhou, G.H. (2007). Effects of high pressure/thermal
treatment on lipid oxidation in beef and chicken muscle. Food Chem. 104: 1575–1579.
Ma, L., and Su, Y.C. (2011). Validation of high pressure processing for inactivating Vibrio parahaemolyticus
in Pacific oysters (Crassostrea gigas). Int J Food Microbiol. 144: 469–474.
Marcos, B., Aymerich, T., and Garriga, M. (2005). Evaluation of high pressure processing as an additional
hurdle to control Listeria monocytogenes and Salmonella enterica in low-acid fermented sausages.
J Food Sci. 70: 339–344.
Marcos, B., Aymerich, T., Monfort, J.M., and Garriga, M. (2008). High-pressure processing and antimicro-
bial biodegradable packaging to control Listeria monocytogenes during storage of cooked ham. Food
Microbiol. 25: 177–182.
Marcos, B., Kerry, J.P., and Mullen, A.M. (2010). High-pressure-induced changes on sarcoplasmic protein
fraction and quality indicators. Meat Sci. 85: 115–120.
Mariutti, L.R., Orlien, V., Bragagnolo, N., and Skibsted, L.H. (2008). Effect of sage and garlic on lipid oxida-
tion in high-pressure processed chicken meat. Eur Food Res Technol. 227: 337–344.
McArdle, R., Marcos, B., Kerry, J.P., and Mullen, A. (2010). Monitoring the effects of high pressure process-
ing and temperature on selected beef quality attributes. Meat Sci. 86: 629–634.
Montiel, R., Alba, M., Bravo, D., Gaya, P., and Medina, M. (2012). Effect of high pressure treatments on
smoked cod quality during refrigerated storage. Food Cont. 23: 429–436.
Mor-Mur, M., and Yuste, J. (2003). High pressure processing applied to cooked sausage manufacture: Physical
properties and sensory analysis. Meat Sci. 65: 1187–1191.
Moussa, M., Perrier-Cornet, J.M., and Gervais, P. (2007). Damage in Escherichia coli cells treated with a com-
bination of high hydrostatic pressure and subzero temperature. Appl Environ Microbiol. 73: 6508–6518.
Murchie, L.W., Cruz-Romero, M., Kerry, J.P., Linton, M., Patterson, M.F., Smiddy, M., and Kelly, A.L. (2005).
High pressure processing of shellfish: A review of microbiological and other quality aspects. Innov
Nishiwaki, T., Ikeuchi, Y., and Suzuki, A. (1996). Effects of high pressure treatment on Mg enhanced ATPase
activity of rabbit myofibrils. Meat Sci. 43: 145–155.
Ohmori, T., Shigehisa, T., Taji, S., and Hayashi, R. (1991). Effect of high-pressure on the protease activity in
meat. Agric Biol Chem. 55: 357–361.
Ohshima, T., Ushio, H., and Koizumi, C. (1993). High-pressure processing of fish and fish products. Trends
Food Sci Technol. 4: 370–375.
Orlien, V., Hansen, E., and Skibsted, L.H. (2000). Lipid oxidation in high-pressure processed chicken breast
muscle during chill storage: Critical working pressure in relation to oxidation mechanism. Eur Food
Res Technol. 211: 99–104.
Patterson, M.F. (2005). Microbiology of pressure‐treated foods. J Appl Microbiol. 98(6): 1400–1409.
Rademacher, B. (2006). Ultrahochdruckverfahren zur Keiminaktivierung. Chem Ing Tech. 78: 1674–1681.
Rastogi, N.K., Raghavarao, K.S.M.S., Balasubramaniam, V.M., Niranjan, K., and Knorr, D. (2007).
Opportunities and challenges in high pressure processing of foods. Crit Rev Food Sci Nutr. 47:
69–112.
Reddy, K.J., Jayathilakan, K., Chauhan, O.P., Pandey, M.C., and Radhakrishna, K. (2015). Effect of high-
pressure processing on physico-chemical and microbial quality characteristics of Chevon (Capra
aegagrus hircus). Food Bioprocess Technol. 8: 2347–2358.
Rendueles, E., Omer, M.K., Alvseike, O., Alonso-Calleja, C., Capita, R., and Prieto, M. (2011). Microbiological
food safety assessment of high hydrostatic pressure processing: A review. LWT-Food Sci Technol. 44:
1251–1260.
Rivas-Cañedo, A., Fernandez-Garcia, E., and Nunez, M. (2009). Volatile compounds in fresh meats subjected
to high pressure processing: Effect of the packaging material. Meat Sci. 81: 321–328.
Rodríguez-Calleja, J.M., Cruz-Romero, M.C., O’Sullivan, M.G., García-López, M.L., and Kerry, J.P. (2012).
High-pressure-based hurdle strategy to extend the shelf-life of fresh chicken breast fillets. Food Cont. 25:
516–524.
Rubio, B., Martinez, B., Garcia-Cachan, M.D., Rovira, J., and Jaime, I. (2007). Effect of high pressure
preservation on the quality of dry cured beef “Cecina de Leon”. Innov Food Sci Emerg Technol. 8:
102–110.
Ruiz-Capillas, C., Carballo, J., and Jiménez-Colmenero, F. (2007b). Consequences of high-pressure process-
ing of vacuum-packaged frankfurters on the formation of polyamines: Effect of chilled storage. Food
Chem. 104: 202–208.
Ruiz-Capillas, C., Colmenero, F.J., Carrascosa, A.V., and Muñoz, R. (2007a). Biogenic amine production in
Spanish dry-cured “chorizo” sausage treated with high-pressure and kept in chilled storage. Meat Sci.
77: 365–371.
Rusman, H., Gerelt, B., Yamamoto, S., Nishiumi, T., and Suzuki, A. (2007). Combined effects of high pressure
and heat on shear value and histological characteristics of bovine skeletal muscle. Asian- Australasian
J Animal Sci. 20: 994–1001.
Sayd, T., Morzel, M., Chambon, C., Franck, M., Figwer, P., Larzul, C., and Laville, E. (2006). Proteome analy-
sis of the sarcoplasmic fraction of pig semi-membranous muscle: Implications on meat colour develop-
ment. J Agric Food Chem. 54: 2732–2737.
Schindler, S., Krings, U., Berger, R.G., and Orlien, V. (2010). Aroma development in high pressure treated beef
and chicken meat compared to raw and heat treated. Meat Sci. 86: 317–323.
Schmidgall, V.J., Toepfl, S., Hertel, C., Bindrich, U., and Heinz, V. (2011). High hydrostatic pressure treatment
of marinated turkey Part 2: Improvement of product safety and production planning. Fleisch. 91: 85–91.
Shigehisa, T., Ohmori, T., Saito, A., Taji, S., and Hayashi, R. (1991). Effects of high hydrostatic pressure on
characteristics of pork slurries and inactivation of microorganisms associated with meat and meat prod-
ucts. Int J Food Microbiol. 12: 207–215.
Sikes, A.L., Tobin, A.B., and Tume, R.K. (2009). Use of high pressure to reduce cook loss and improve texture
of low-salt beef sausage batters. Innov Food Sci Emerg Technol. 10: 405–412.
Simonin, H., Duranton, F., and de Lamballerie, M. (2012). New insights into the high-pressure processing of
meat and meat products. Compr Rev Food Sci Food Safety. 11: 285–306.
Simpson, R.K., and Gilmour, A. (1997). The effect of high hydrostatic pressure on Listeria monocytogenes in
phosphate‐buffered saline and model food systems. J Appl Microbiol. 83: 181–188.
Smelt, J.P.P.M. (1998). Recent advances in the microbiology of high pressure processing. Trends Food Sci
Technol. 9: 152–158.
Souza, C.M., Boler, D.D., Clark, D.L., Kutzler, L.W., Holmer, S.F., Summerfield, J.W., and Killefer, J. (2011).
The effects of high pressure processing on pork quality, palatability, and further processed products.
Meat Sci. 87: 419–427.
Sun, D.W., and Norton, T. (2008). Recent advances in the use of high pressure as an effective processing tech-
nique in the food industry. Food Bioprocess Technol. 1: 2–34.
Sun, X.D., and Holley, R.A. (2010). High hydrostatic pressure effects on the texture of meat and meat products.
J Food Sci. 75: R17–R23.
Suzuki, A., Watanabe, M., Iwamura, K., Ikeuchi, Y., and Saito, M. (1990). Effect of high pressure treatment
on the ultrastructure and myofibrillar protein of beef skeletal muscle (food and nutrition). Agric Biol
Chem. 54: 3085–3091.
Szerman, N., Barrio, Y., Schroeder, B., Martinez, P., Sancho, A.M., Sanow, C., and Vaudagna, S.R. (2011).
Effect of high hydrostatic pressure treatments on physico-chemical properties, microbial quality and
sensory attributes of beef carpaccio. Procedia Food Sci. 1: 854–861.
Tintchev, F., Bindrich, U., Toepfl, S., Strijowski, U., Heinz, V., and Knorr, D. (2013). High hydrostatic pres-
sure/temperature modelling of frankfurter batters. Meat Sci. 94: 376–387.
Tintchev, F., Wackerbarth, H., Kuhlmann, U., Toepfl, S., Knorr, D., Hildebrandt, P., and Heinz, V. (2010).
Molecular effects of high-pressure processing on food studied by resonance Raman. Annals New York
Academy Sci. 1189: 34–42.
Toepfl, S., and Heinz, V. (2009). New options for targeted product modification. Fleisch Int. 3: 11–13.
Toepfl, S., Mathys, A., Heinz, V., and Knorr, D. (2006). Review: Potential of emerging technologies for energy
efficient and environmentally friendly food processing. Food Rev Int. 22: 405–423.
Tornberg, E. (2005). Effects of heat on meat proteins–Implications on structure and quality of meat products.
Meat Sci. 70: 493–508.
Tornberg, E., Andersson, K., and Josell, A. (1997). The rheological properties of whole and minced meat dur-
ing cooking as related to sensory and structural characteristics. In: Proceedings of the 1st International
Symposium on Food Rheology and Structure, pp. 16–20/3, Zurich.
Trespalacios, P., and Pla, R. (2007a). Simultaneous application of transglutaminase and high pressure to
improve functional properties of chicken meat gels. Food Chem. 100: 264–272.
Trespalacios, P., and Pla, R. (2007b). Synergistic action of transglutaminase and high pressure on chicken
meat and egg gels in absence of phosphates. Food Chem. 104: 1718–1727.
Tume, R.K., Sikes, A.L., and Smith, S.B. (2010). Enriching M. Sternomandibularis muscle with alpha-tocoph-
erol by dietary means does not protect against the lipid oxidation caused by high-pressure processing.
Meat Sci. 84(1): 66–70.
Wackerbarth, H., Kuhlmann, U., Tintchev, F., Heinz, V., and Hildebrandt, P. (2009). Structural changes of
myoglobin in pressure-treated pork meat probed by resonance Raman spectroscopy. Food Chem. 115:
1194–1198.
Wiggers, S.B., Kroger-Ohlsen, M.V., and Skibsted, L.H. (2004). Lipid oxidation in high-pressure processed
chicken breast during chill storage and subsequent heat treatment: Effect of working pressure, packag-
ing atmosphere and storage time. Eur Food Res Technol. 219: 167–170.
Wood, J.D., Enser, M., Fisher, A.V., Nute, G.R., Sheard, P.R., Richardson, R.I., and Whittington, F.M. (2008).
Fat deposition, fatty acid composition and meat quality: A review. Meat Sci. 78: 343–358.
Yagiz, Y., Kristinsson, H.G., Balaban, M.O., Welt, B.A., Ralat, M., and Marshall, M.R. (2009). Effect of high
pressure processing and cooking treatment on the quality of Atlantic salmon. Food Chem. 116: 828–835.
Yang, H., Han, M., Bai, Y., Han, Y., Xu, X., and Zhou, G. (2015). High pressure processing alters water dis-
tribution enabling the production of reduced-fat and reduced-salt pork sausages. Meat Sci. 102: 69–78.
Yoshioka, K., Kage, Y., and Omura, H. (1992). Effect of high pressure on texture and ultrastructure of fish and
chicken muscles and their gels. In: High Pressure and Bioscience, pp. 325–327. Balny, C., Hayashi, R.,
Heremans, K., and Masson, P., Eds. John Libbey Eurotext, Montrouge, France.
Yuste, J., Pla, R., Capellas, M., Sendra, E., Beltran, E., and Mor‐Mur, M. (2001). Oscillatory high pressure pro-
cessing applied to mechanically recovered poultry meat for bacterial inactivation. J Food Sci. 66: 482–484.
ChaptEr 5
high-pressure processing of Milk and Milk products
Ashish Kumar Singh, Sanket Borad, G. S. Meena, Heena Sharma, and Sumit Arora
CONtENtS
5.1 Introduction ............................................................................................................................ 69

5.2 Principle of High-Pressure Processing ................................................................................... 70
5.3 Effect of HHP on Milk Microorganisms ................................................................................ 71
5.4 High-Pressure-Induced Effects on Milk Constituents............................................................ 74
5.4.1 High-Pressure-Induced Effect on Water and Minerals .............................................. 75
5.4.2 High-Pressure-Induced Effects on Milk Proteins ...................................................... 75
5.4.3 Effect of High-Pressure Treatment on Milk Fat, Lactose, and Micronutrients .......... 78
5.5 High-Pressure-Mediated Effect on Milk Enzymes ................................................................ 78
5.6 High-Pressure-Related Changes on Functional Characteristics of Milk ............................... 79
5.7 Prospective Applications of High-Pressure Processing in Dairy Industry ............................80
5.7.1 HHP Induced Effect on Cheese Making Characteristics ...........................................80
5.7.2 Yoghurt and Ice-Cream .............................................................................................. 82
5.7.3 Prospective Applications in Functional Dairy Foods ................................................. 82
5.8 Conclusion .............................................................................................................................. 83
References ........................................................................................................................................84
5.1 INtrODUCtION
Among the dietary components, milk and milk nutrients occupy a special place. Consumption
of liquid milk and other dairy products is increasing globally. Milk is served as ideal base material
for the production of value-added products by applying various processing technologies such as
thermal processing, freezing, fermentation, concentration, and fractionation. The richness of milk
in a wide array of bioactives has also generated interest among stakeholders, including processors,
consumers, dieticians, medical practitioners, and marketing professionals. Milk was among the few
commodities on which the first experimentation related to high-pressure processing (HPP) was car-
ried out almost a century ago by Hite (1899) at Virginia Experimental Station. Despite holding the
promise to inactivate the microorganisms associated with milk spoilage, HHP could not be further
utilized in milk and other food products owing to various issues related to equipment design and
their operation on a commercial scale. However, research carried out recently indicate that HHP
seems a very promising processing intervention, as it offers numerous opportunities for developing
shelf-stable foods with better nutrition, excellent organoleptic characteristics, and in delivering safe
foods to consumers (Fonberg-Broczek et al., 1999). The technology has potential to address certain
69
critical issues associated with conventional thermal processing in milk and milk products such
as browning, loss of nutrients, and bioactive components. Since the temperature employed in the
majority of the HHP processes is in the ambient range, it may potentially reduce energy consump-
tion and costs of operation, as well as improve sustainability of production (Toepfl et al., 2006). The
most attractive feature of HHP, which has made the process worldwide acceptable, is uniform pro-
cessing ability as the pressure is applied uniformly throughout the food material, independent of its
mass and time. In the case of dairy products, high pressure (up to 400 MPa) has been used to process
milk, which has successfully yield desirable characteristics of milk products like yogurt, cheese,
and cream, among others; in fact, there have been more studies on these products than on fluid milk
alone. Besides inactivation of microorganisms and shelf life extension, HHP also homogenizes milk
(O’Reilly et al., 2001), which is similar to milk processed under ultra-high temperature conditions.
For low-acid foods, such as milk, mild temperature in combination with high pressure is necessary
to ensure extended shelf life products with better quality characteristics (Balasubramaniam and
Balasubramanian, 2003). Pressure in combination with heat can be used for sterilization of milk.
Researchers have shown that the application of ultra-high pressure (UHP) combined with mild tem-
peratures can extend the shelf life of milk up to 45 days.
5.2 prINCIpLE OF hIGh-prESSUrE prOCESSING
The HPP working principle is based on two important laws. The basic thermodynamics rule
explaining the application of high pressure in food processing operations is based on Le Chatelier’s
principle. According to this, any reaction of phase transition phenomenon accompanied by reduc-
tion in volume is favoured by high pressure, whereas those leading to volume expansion would be
inhibited. On the other hand, transition state theory (TST) postulates that the rate constant of any
reaction in aqueous phase is proportional to the quasi-equilibrium (chemical equilibrium between
reactants and activated transition state complex). The activated complex can be converted into prod-
ucts and kinetic theory may be used to calculate the rate of the conversion. The driving force in any
reaction is a negative change in free energy (−ΔG) and such reactions are termed as spontaneous
reactions. The basic equation of Gibbs correlates G with pressure, temperature, and chemical poten-
tial. It is well understood that an increase in pressure influences the rate of chemical reactions, but to
a lesser extent than temperature. A pressure of 100 MPa magnitudes would produce a similar effect
as a rise in 10°C in a solution (Tauscher, 1995). The majority of biochemical reactions result in
alteration in volume, hence such reactions are affected by pressure treatments. Food molecules con-
sist of water, proteins, starch, lipids, mineral salts, vitamins, and a large number of other molecules.
High-pressure treatment exerts a multitude of effects of different magnitude on these molecules,
thereby influencing the characteristics of food products.
Another important feature of high-pressure treatment is its isostatic nature; if pressure is
applied to food materials, it will immediately be transmitted across the food uniformly indepen-
dent of its size shape, geometry, and dimensions. A typical HPP system consists of a treatment
chamber, a pressure generating system, pressure transmission medium, and pressure intensifier.
The treatment chamber is made of high-tensile-strength stainless steel alloy or other suitable con-
struction material. One of the limitations in construction of HP systems is non-availability of suit-
able fabrication material. In batch-type systems, the food material, whether liquid or semi-solid or
solid, is packaged in high strength flexible packaging material, packaging is degassed to remove
air and immersed in a pressurization chamber/vessel, the vessel is closed, and then it is subjected
to high-pressure treatment using high quality water or glycerol or combination thereof as pres-
surizing medium for the specified period. The mineral oil or vegetable oil is often employed as
anti-corrosive material. After holding for the requisite period, the vessel is opened and the package
is removed from it and kept normally under cold chain. The equipment is also having provisions
HiGH-Pressure ProCessinG oF MiLK AnD MiLK ProDuCts 71
for monitoring the process variables and control devices for temperature and pressure. Batch-type
systems are normally used for HP treatment of solid or semi-solid food materials, whereas semi-
continuous or continuous systems are preferred for the treatment of viscous and flowable liq-
uid foods, which are pumped through the treatment chambers and held under pressure for the
desired duration. The commonly employed packaging materials for high-pressure-processed foods
include ethylene vinyl and polyvinyl films. Although HPP appears quite simple in terms of plant,
machinery, and operational features, developing a fool-proof system remains a major challenge.
Currently available commercial HP processing equipment can operate in the 100–1000 MPa pres-
sure range. Equipment that can work at slightly higher operational pressure, i.e., 1000–2000 MPa,
is still a daunting task.
5.3 EFFECt OF hhp ON MILK MICrOOrGaNISMS
Being a preservation technology, HPP aimed at inactivation of microbial groups that are involved
in pathogenicity and spoilage of foods. The major site for high pressure-related anti-microbial effects
is microbial membrane, which usually gets ruptured on applying pressure; however, such effect is
more visible above 400 MPa. HP causes a number of morphological and biochemical changes apart
from affecting the cell membrane and genetic material. The lethal effect of HP is mainly attributed
to its effect on cell membrane permeability and also on the activity of membrane-bound ATPase.
Observation of microbial cells under microscope on pressurization indicated loss of ability to move,
elongation in microbial cells, separation of the cell wall with the cell membrane, and structural
deformation, if held for long duration. Cell membrane composed of lipid-protein-lipid bilayer and
integrity of cell membrane play vital roles in its metabolisms and viability. The cell membrane is
semi-permeable in nature, i.e., it permits the entry or removal of cellular components or metabolites.
Lipid bilayer contained embedded functional proteins, i.e., membrane bound enzymes, which are
crucial for the normal metabolic reaction of microbes.
On high-pressure treatment, the lipid layer undergoes phase transmission and becomes solidified,
which adversely affects the fluid nature of the membrane. With increasing pressure, the cell membrane
gets destroyed, resulting in leaking of cellular components. Damage to cell membrane also influence
transport phenomenon involving the nutrient uptake and removal of toxic waste generated. Likewise,
conformational changes in functional proteins severely alter the cellular metabolic reaction including
cell division. The resistance of microorganisms to pressure on food is quite variable depending on
HP processing conditions (magnitude of pressure, period of pressure treatment, temperature, mode
of pressurization), food constituents (moisture content, protein, lipid components, etc.) and the char-
acteristics and the physiological state of the microorganism (Smelt, 1998).
Since, among the microbial groups, bacteria are the most common contaminants and pos-
ing safety threats. Bacterial cells are injured or killed by high hydrostatic pressure (HHP),
depending on pressure levels, species and strain of the microorganism, and subsequent stor-
age conditions. Most of the reports indicate that a pressure treatment in excess of 200 MPa at
ambient temperature is required for bacterial cell inactivation. Normally pressure treatment in
the range of 400–600 MPa destroys most of the vegetative cells of bacteria, yeast, and fungi
at room temperature, but still several pressure-resistant strains may survive under such condi-
tions (Benito et al., 1999). It has also been noticed that injured bacteria may get repaired during
storage, which could be an important safety issue especially in low acid food products. The
pressure resistance of microorganisms can be affected by many intrinsic and environmental
parameters. Gram-positive microorganisms are more resistant to pressure than gram-negative
bacteria because of the presence of teichoic acid in the cell wall of the former. Teichoic acid
content offers more structural integrity and rigidity to bacterial cell wall. Gram-positive micro-
organisms need an application of 500–600 MPa at 25°C for 10 min to achieve inactivation,
while gram-negative microorganisms can be inactivated with treatments of 300–400 MPa with
the same time/temperature combination. Several workers have observed that baro-resistance of
yeast and moulds is lower than bacteria. Vegetative forms of yeasts and moulds are most pres-
sure sensitive compared to spores produced thereof (Smelt, 1998). Microbial cells in logarithmic
growth phase are more baro-sensitive than their stationary phase because of immaturity of cells
and higher metabolic rate, which render them prone to stress.
The nutrient density of a food may influence the microbial cell destruction using high pressure;
probably due to the availability of nutrients microbial cells rejuvenate themselves rapidly. Despite
advancements in understanding of the mechanism of microbial inactivation using high pressure,
it is still not possible to produce a sterilized product by applying HPP alone below pressure
treatment of 1000 MPa. Spores, produced as a strategic intervention to overcome adverse envi-
ronmental conditions or stress, are more difficult to inactivate by the majority of processing treat-
ments including thermal and high-pressure processing. In food processing operations, Bacillus
and Clostridia are the two bacterial classes that are of concern owing to their tendency to undergo
sporulation. These two groups have been reported in the past with numerous spoilage and food-
borne illness incidences and the problem could be more severe under anaerobic conditions existing
inside the package. Lower pressure treatment remains ineffective in inactivating the spores and
normally the bacterial spores can survive at a pressure of 1000 MPa (Cheftel, 1995). However, it
has been noticed that the pressurization along with mild heat treatment triggers spores to germi-
nate and after germination, microorganisms lose their resistance towards pressure and heat, and
get killed (Gould and Sale, 1970, 1988; Knorr, 1995; Gould, 2000). Acidothermophilic bacteria
like Alicyclobacillusacidoterrestris and Bacillus coagulans can cause spoilage of heat-processed
acidic foods because they form spores with very high heat resistance and can grow at low pH
(Vercammen et al., 2012). The groups investigated the germination and inactivation of spores
of two bacterial groups in the buffer and tomato sauce using high pressurization and reported
that spore germination in the low-pressure zone was not much influenced by temperature; how-
ever, strong temperature dependence was noted at high pressure range. They further concluded
that mild heating at 60°C may induce spore germination, which can be subsequently killed by
high-pressure treatments. Thus, pH of the medium also has an effect on spore germination and
its inactivation. Among the other approaches attempted are thermization followed by pressure
treatment and low pressure-induced germination followed by high-pressure or temperature treat-
ment and application of bio-preservatives (Nisin and bacteriocin) along with HPP. However, fur-
ther systemic experimentations with diverse microbial groups is necessary before suggesting a
fool-proof treatment.
Milk serves as an ideal growth medium for microbial proliferation, both spoilage and patho-
genic ones. Milk gets contaminated by a wide range of microbial groups once it comes out of the
udder and the nature of contaminating microorganisms depends on the health status of animal,
hygiene of animal, workers and environment around it, temperature prevailing, and extent of
clean milk production practices followed. Typical milk microbiota include aerobe psychrotrophic
gram-negative bacteria, yeast, fungi, heterofermentative lactobacilli and spore-forming bacteria
(Ledenbach and Marshall, 2009). Approximately 65%–70% of psychrotrophs isolated from raw
milk samples belongs to Pseudomonas spp. and these are known to produce heat-resistant proteo-
lytic and lipolytic enzymes. Other psychrotrophic groups associated with raw milk spoilage are
Micrococcus, Aerococcus, Lactococcus, Bacillus, and members of Enterobacteriaceae. Coliforms,
lactic acid bacteria, fungi, yeast, and spore former bacteria (Bacillus and Clostridia) may also
be associated with spoilage of raw milk and processed dairy products. Chilling of raw milk is
normally practiced to check the proliferation of contaminating microbes. However, after 48 h
storage under chilled conditions the population of most spoilage bacteria, especially psychrotro-
phic Pseudomonas, grows rapidly leading to faster deterioration of raw milk quality and spoilage.
Thermal treatments like thermization, pasteurization, and UHT processing are applied to extend
the shelf-life and storage stability of liquid milk. However, heating of milk may adversely affect
the sensory, nutritional, and other functional characteristics of milk. An alternative of thermal
treatment has always been sought to overcome the problems associated with conventional thermal
processing of milk.
In the recent past, presence of pathogenic microorganisms such as Listeria spp., Salmonella spp.,
Shigella spp., Yerciniaenterocolitica, and an acid-resistant pathogenic strain of E. coli, i.e., E. coli
O157:H7, in dairy products has raised safety concerns. Moreover, emergence of zoonotic diseases
raised concern of microbial safety of raw and processed dairy products. Many studies have been
conducted on raw milk using high pressure and have proved that HPP treatment gives raw milk
quality (pressurized at 400–600 MPa) comparable to that of pasteurized milk (but not that of steril-
ized milk due to presence of HP-resistant spores), as it is equally effective in destroying pathogenic
and spoilage microorganisms. To achieve a shelf life of 10 days at 10°C storage, a pressure treatment
of 400 MPa for 15 min or 600 MPa for 3 min at 20°C is necessary (Rademacher and Kessler, 1997).
In a study, pressurization of samples was carried out using a high pressure at 41°C and 448 MPa for
11 min. Treatment of L. monocytogenes in milk under HPP was not as effective as in fruit juices,
because the high nutrient content in whole milk increased the resistance of the strain to HPP. HPP
was suitable for inactivation of microorganisms in high- and low-acid food systems as compared
to food with higher pH as in the case of milk. Studies conducted by Vachon et al. (2002) revealed
that dynamic high-pressure treatment inactivates the three major food pathogens including Listeria
monocytogenes (LSD 105-1), Escherichia Coli O157:H7 (ATCC 35150), and Salmonella enteritidis
(ATCC 13047) present in milk. In a similar experimentation carried out by Gervilla et al. (2000),
milk inoculated with five microbial groups like Escherichia coli CECT 405 (indicator of hygiene
conditions), Pseudomonas fluorescens CECT 378 (representative of major microbial group present
in chilled milk), Listeria innocua CECT 910 (indicator of human-pathogen L. monocytogenes),
Staphylococcus aureus CECT 534 (major microbial group associated with mastitic milks), and
Lactobacillus helveticus CECT 414 (representative of starter lactic acid bacteria present in fer-
mented milk) was processed using high pressure. The microbial inactivation followed the sequence
of Pseudomonas fluorescens > Escherichia coli > Listeria innocua > Lactobacillus helveticus >
Staphylococcus aureus. However, the treatment temperature had a different effect: P. Fluorescens,
L. innocua, and L. helveticus were more resistant at 25°C than 4°C, while the reverse was observed
for E. coli and S. aureus.
Mussa and Ramaswamy (1997) evaluated the UHP requirements of raw milk for shelf-life
enhancement. They applied the UHP in the range of 200–400 MPa for the period ranging from
5 to 120 min and kinetic parameters like rate constant and decimal reduction time were com-
puted. Kinetic parameters followed the first-order reaction and as expected increasing the pres-
surization caused higher mortality and inactivation of enzymes. The pressure z values (zp) for
the same variables were 168, 398, 404, and 532 MPa, respectively. Milk subjected to a microbial
4D UHP process at 350 MPa had a shelf-life of 25 days at 0°C, 18 days at 5°C, and 12 days at
10°C, respectively. Hayman et al. (2008) observed that lowering water activity (aw) of the system
increased the resistance of microorganisms to high pressures; however, it depends on the solute
used to depress aw. The sharp increase in barotolerance of L. monocytogenes below aw 0.83 may
be due to protein stability, because at lower water activity proteins are inflexible and hence
did not get denatured by HPP treatment. Therefore, the bacterial cells were not inactivated by
pressure. The strong correlations between aw and bacterial survival (r2 = 0.84) and aw and LDH
activity (r2 = 0.98) indicated a relationship exists between protein denaturation and bacterial
inactivation during HPP. It can be inferred that microbial inactivation by applying high pres-
sure is influenced by several factors and process conditions need to be optimized to completely
eliminate the microorganisms.
5.4 hIGh-prESSUrE-INDUCED EFFECtS ON MILK CONStItUENtS
Milk is a colloidal suspension of casein micelles, in which milk lipids are dispersed in the
form of emulsion and other solutes including lactose, serum proteins, minerals, and other water-
soluble constituents are dispersed in solution phase. Bovine milk contains about 87% moisture,
3.5%–4.0% fat, 3.5% protein, 4.5% lactose, and 0.8%–1.0% ash content. The composition of milk
depends on several factors such as breed, stage, and number of lactations, seasons, feeding prac-
tices, and animal management practices followed. Among the milk proteins casein constitutes
80% and the remaining 20% are whey proteins. Milk constituents are altered by various food
processing technologies including thermal treatments, freezing, concentration, evaporation,
acidification, and irradiation, which may have positive or negative influence on their functional
properties, sensory characteristics, and nutritive values. HPP, though a non-thermal technology,
has been observed to modify the inherent features of milk constituents and hence substantially
change the properties of treated milk. The high pressure-induced effects on milk constituents
have been reviewed by a number of researchers previously and excellent information is criti-
cally examined by them (Huppertz et al., 2002, 2006a; Trujillo et al. 2002). High pressure led
to modifications in milk constituents of varying degrees depending on its level and period of
pressurization (Table 5.1).
table 5.1 high-pressure-Induced Effects on Milk Constituents

Milk Constituents Effects of hhp references
Casein extensive disruption of casein micelles in the Harte et al. (2007)
250–310 MPa range. Addition of whey protein to
casein isolates protected the micelles from
high-pressure-induced disruption
increase in size and number of casein micelles Huppertz et al. (2006b)
Whey proteins Denaturation of whey proteins Chicon et al. (2008)
enhances pepsin hydrolysis of β-lactoglobulin
(β-lg) at 400 MPa
reduction in antigenicity and immunoglobulin e
(ige) binding of β-lg
improving functional properties of whey proteins Liu et al. (2005b)
Fat HHP up to 500 MPa catalyses modifications in Gervilla et al. (2001)
size and distribution of milk fat globules
Crystallization of milk fat in cream at HP treatment buchheim and Abou el-nour (1992)
at 100–400 MPa and maximum crystallization
occurred at 200 MPa
At up to 200 MPa, the crystallization and melting Frede and buchheim (2000)
temperatures of milk fat are increased by 16.31°C
and 15.51°C/100 MPa, respectively
Lactose no changes have been observed after Lopez-Fandino et al. (1996)
pressurization (100–400 MPa for 10–60 min at
25°C)
Flavour HHP at 600 MPa at 50°C resulted in an increase Liu et al. (2005a)
in number of flavour-binding sites of WPC
Water Decrease in its freezing point, to −4, −8 or −21°C Hinrichs et al. (1996a)
at 50, 100 or 210 MPa, respectively
Compressed considerably by about 4% at
100 MPa or 15% at 600 MPa
Minerals HP treatment solubilized both indigenous and buchheim (1996); schrader et al.
heat-precipitated CCP (1997)
5.4.1 high-pressure-Induced Effect on Water and Minerals
Water, being an important constituent for the majority of dairy products, has influence on their
quality characteristics and stability. In dairy products, water molecules mainly act as solvent and are
determinant for large number of functionalities such as solubility, gelation, emulsification, disper-
sion, aeration, foaming, and film formation of macromolecules. On pressurization, water molecules
get compressed to varying extents depending on the amount of pressure. Hinrichs et al. (1996a)
reported 4% and 15% compression at 100 and 500 MPa, respectively. They further observed linear
lowering in the freezing point of water with extent of pressurization. Ice formation is accompanied
by increase in volume and generation of heat; hence, both cause decrease in freezing point. This
phenomenon could be useful in preventing freezing-induced injuries or defects in frozen raw and
processed foods especially during storage. High pressure led to dissociation of water molecule lead-
ing to decrease in pH. With 1000 MPa increase in pressure, the pH of water is lowered by one unit.
Effects of HP on minerals in milk can be divided into (1) effects on the distribution between the
colloidal and diffusible phases and (2) effects on ionisation. HP treatment at 400 MPa of previously
pasteurized or high temperature-treated milk increased diffusible calcium levels indicating that the
HP treatment solubilised both indigenous and heat-precipitated colloidal calcium phosphate (CCP)
(Buchheim et al., 1996; Schrader et al., 1997). Lopez-Fandino et al. (1998) reported that the con-
centrations of calcium, magnesium, and phosphorus in the diffusible phase of bovine or caprine
milk increased following treatment up to 300 MPa, but subsequently decreased when magnitude of
pressure was enhanced at 400 MPa. HP-induced changes in the level of diffusible salts were more
pronounced in ovine milk and increased with pressure up to 400 MPa (Lopez-Fandino et al., 1998).
In contrast, Law et al. (1998) noted little effect of treatment at 100–500 MPa at 20°C or 45°C on
the levels of colloidal calcium and phosphorus in caprine milk. There could be various possible
underlying mechanisms that can be suggested for the variation in observations. Processing-related
alterations in milk minerals have been associated with colour, thermal stability, and rennet coagula-
tion properties of milk.
5.4.2 high-pressure-Induced Effects on Milk proteins
Messens et al. (1997) reviewed the literature pertaining to high pressure-induced changes in
food proteins and its effect on their functionality. They suggested that proteins in native state are
stabilized by covalent bonds (including disulphide bridges) plus electrostatic interactions (ion
pairs, polar groups), hydrogen bridges, and hydrophobic interactions. Covalent bonds are almost
unaffected by HP and hence the primary structure of proteins remains intact during HP treatment.
On the other hand, changes in secondary structure occur at high pressures and lead to irreversible
denaturation, because stabilizing hydrogen bonds are enhanced at low pressures and ruptured at
very high pressure. Significant changes in the tertiary structure of proteins, which are maintained
mainly by hydrophobic and ionic interactions, are observed above 200 MPa. Ionic bonds in aque-
ous solutions are strongly destabilized by pressure as in the vicinity of each ion, water molecules
are arranged more densely than in bulk water, leading to decrease in volume. Hydrophobic interac-
tions are characterized by increase in volume and are hence destabilized at elevated pressure.
Multimeric proteins, held together by non-covalent bonds, thereby disrupt quaternary structure.
On the basis of screening of scientific investigations on high pressure-related conformational
changes in food proteins, they concluded that such changes may lead to protein denaturation,
aggregation, and gelation depending on protein properties (type of protein, pH, ionic strength,
water content) and amount and duration of pressure applied. Such modifications in functional
properties of food proteins including milk proteins can be exploited to improve the textural attri-
butes of dairy products, enhancement of functionality of milk protein ingredients, and develop-
ment of a new range of food products. Caseins are phosphoprotein with an average molecular
weight of about 20 KDa, which consist of four proteins, i.e., αs1 (34%–40%), αs2 (11%–15%),
β-casein (25%–35%), and κ-casein (8%–15%). The hydrophobic core usually contains αs1, αs2 and
β-casein, while peripheral hydrophilic portion contains κ-casein. Casein particles on an average
contain 94% proteins and 4% mineral matter on dry weight basis. Casein particles are present in an
associated form referred to as micellar structure with micelle size varying in the range of
50–300 nm. Micelles are a highly hydrated form, normally containing 3.3 g of water per gram of
protein. Micelles are quite porous and occupy 4 mL/g. Caseins have low level of secondary and
tertiary structures and 90% of it exists in the form of macromolecular aggregates. Several models
have been proposed for microstructure of casein; however, it is widely accepted that the framework
of the casein micelles is formed by the nanoclusters/nanotubes, which comprise an amorphous
MCP core with a radius of approximately 2–3 nm, surrounded by a shell of phosphopeptides
(caseins), which contribute phosphoseryl clusters (Holt et al., 1998). These nanoclusters are distrib-
uted almost homogeneously throughout the micelles and are joined through hydrophobic bonding
and electrostatic interactions. κ-Casein protruding outwardly from the surface offers electrostatic
and steric repulsion, thus checking inter-micellar aggregation. In HPP, any change associated with
volume reduction is favoured by high pressure. High pressure influences the properties of milk and
milk components; however, the effect may depend on native structure of macromolecules and the
extent of pressurization. HP treatment does not affect covalent bonds in the temperature range of
0°C–40°C, so the primary structure of protein remains intact on pressurization; however, it influ-
ences electrostatic an ionic interaction responsible for the maintenance of secondary structure. At
pressure above 200 MPa significant changes in tertiary structure are observed. The casein micelles
are disintegrated into smaller particles resulting in an increase of caseins and calcium phosphate
levels in the serum phase of milk and a decrease in the both non-casein nitrogen and serum nitro-
gen fractions (Law et al., 1998). Pressure treatment in the range of 100–300 MPa generally tends
to be reversible, but above 300 MPa it leads to irreversible denaturation (Jaenicke, 1981).
Pressurization of milk causes conformational changes in milk proteins and on applying HP treat-
ment the size and number of casein micelles increases, because the spherical particles join together
to form chains or clusters of sub-micelles. Researchers reported no appreciable changes in casein
micelle size on pressure treatment in the range of 150–200 MPa; however, increasing the pressure
level from 150 to 600 MPa resulted in 40%–50% reduction in casein micelle size. Transmission
electron microscopic (TEM) examination revealed that pressure treatment of 400 or 600 MPa
completely disrupted the caseins micelles into smaller fragments (Needs et al., 2000). Dissociation
of caseins is also affected by the pH of milk and the level of caseins was higher at pH 5.5 or
7.0 than the 6.7, the natural pH of milk (Arias et al., 2000). It is attributed to increased dissociation
of colloidal calcium caseinate (CCP) complex or increased electrostatic repulsion. However, the
effect of pressure treatment on casein moiety is temperature dependent as well. Gaucheron et al.
(1997) noticed that at constant pressure (250 MPa), the temperature affects the alternation in casein
micelles and casein micelle size increased at 4°C, remained same at 20°C, but decreased at ele-
vated temperature (40°C). HP treatment increases the transfer of individual caseins from the col-
loidal to the soluble phase and dissociation of the caseins is in the order of κ>β>αs1>αs2. The order
of dissociation of caseins largely corresponds to the serine phosphate content of the caseins, indi-
cating that caseins (which are more tightly bound to CCP) dissociated to a lesser extent (Aoki et al.,
1990). Higher serine-phosphate content reflected in enhanced baro-resistant for the caseins. The
serine phosphate residues are 1, 5, 8, and 11 for κ, β, αs1, and αs2 caseins, respectively. Differential
inactivation of caseins on pressure treatment could be exploited for the fractionation of individual
caseins and harness their unique functional properties. Among the whey proteins β-lactoglobulin
denaturation initiates above 150 MPa, but complete denaturation occurs above 500 MPa at 25°C.
β-Lactoglobulin is the most heat-sensitive milk protein and its denaturation starts at 65°C and
completes at 90°C. A similarity was also observed with high-pressure treatment, wherein pres-
surization at 400 MPa denatured approximately 70%–80% β-lactoglobulin. However, further
increase in level of pressure had little effect on denaturation severity that showed that its denatur-
ation is concentration dependent. The denaturation rate constant order for β-lactoglobulin was 2.5,
further confirming its concentration dependency (Hinrichs and Rademacher, 2004). A synergistic
effect of temperature and pressure on the denaturation of β-lg has been reported. Similar levels of
denaturation (~100%) were noticed on treatment of raw milk at 300 MPa at 60°C or at 400 MPa at
40°C. However, denaturation was reduced by HP treatment at 4°C relative to 20°C. HP-induced
denaturation of β-lg decreases on milk acidification to pH 5.5 or 6.0 before treatment and is
increased at pH 7.0, relative to milk at pH 6.7. Increased denaturation of β-lg at pH 7.0 may be
attributed to enhanced reactivity of its free -SH group at alkaline pH values (Huppertz et al.,
2004a). α-la in bovine milk is more resistant to denaturation under pressure and its denaturation
occurs at pressures above 500 MPa. Similar observations have been reported for ovine and caprine
milk. However, treatment at higher temperatures (50°C–60°C) greatly increased the extent of
denaturation. Differences in the barostability of α-la and β-lg may be linked to the more rigid
molecular structure of the former, caused partially by the numbers of intra-molecular -S-S- in the
two proteins and the lack of free –SH in α-la (Hinrichs et al., 1996b; Huppertz et al., 2004c). High
pressure-induced changes on milk proteins have a marked effect on physico-chemical properties
of milk such as heat stability, viscosity, surface tension, colour, and, ultimately, on functional prop-
erties as well. Borad et al. (2017) compiled the effects of processing on nutritive values of milk
proteins with special emphasis on HHP. The HHP has been shown to improve the nutritive values
of milk proteins by reducing their antigenicity depending on pressure-temperature combinations.
β-Lg has been reported to exhibit an improved proteolytic digestibility when hydrolysed under
HHP. It is suggested that the enhancement of proteolysis under HP conditions is due to conforma-
tional changes of β-Lg that make it more susceptible to hydrolysis, rather than on pressure effects on
the enzymes or direct effect on the β-Lg-enzyme interaction (Dufour et al., 1995; Stapelfeldt et al.,
1996; Bonomi et al., 2003). β-Lg, resisting the proteolytic digestion by pepsin and chymotrypsin
in native conformation, can be hydrolysed slightly by pepsin when treated for HHP treatment at
400 MPa for 10 min. The rate of enzymatic hydrolysis/digestion can be enhanced at elevated pres-
sure treatment of 600 and 800 MPa. Further, the resultant peptides liberated post-hydrolysis had
molecular weight less than 1.5 kDa. Therefore, there exists a great scope for utilization of HHP
treatment for the development of infant formulae as well as dairy products with milk protein aller-
genicity (Zeece et al., 2008). Proteolysis of β-Lg under high pressure involves the rapid disappear-
ance of the protein, leading to the accumulation of intermediate peptides that are further degraded
on extending incubation with the enzyme. The potential of HHP to produce whey protein hydroly-
sates with acceptable functional properties and reduced potential allergenicity for use in hypoal-
lergenic foods was studied by Chicon et al. (2008).
Upon the HHP treatment of 200 and 400 MPa for 15 min, rapid digestion of αs1- and β-caseins
by plasmin was noticed as compared to control. The possible mechanism elucidated was also
explained wherein casein micelles disintegrate and solubilize under the influence of high pressure,
leading to exposing the sites for enzymatic cleavage to proteolysis (Garcia-Risco et al., 2003). HHP
of 100–300 MPa has been shown to enhance overall enzymatic (trypsin, chymotrypsin, and pepsin)
hydrolysis of whey proteins and reduced the residual antigenicity of the whey protein hydrolysates
(Penas et al., 2006). Some contradictory effects of HHP on milk protein allergenicity have also been
reported wherein antigenicity of β-Lg was elevated post-HHP (200–600 MPa) treatments to whey
protein isolate solution, skim milk, and sweet whey. This could be due to unfolding of protein mol-
ecules exposing the epitome having cross reactivity, which otherwise were buried inside the cavity
(Kleber et al., 2007).
Other whey proteins have not been studied to that greater extent, such as β-lactoglobulin and
α-lactalbumin. Bovine serum albumin (BSA) is quite baro-resistant up to 400 MPa in raw milk
and even above 400 MPa very little denaturation occurs. Lopez-Fandino et al. (1996) did not
observe denaturation of BSA in bovine milk at 100–400 MPa. The high barostability of BSA is
probably related to the fact that this molecule, through its 17 intra-molecular disulphide bonds, has
an extremely rigid molecular structure which remains largely unaffected under HP. Immunoglobulin,
α-Lactalbumin and bovine serum albumins are more resistant and their denaturation occurs at the
highest pressures and at temperature above 50°C. Immunoglobulins in milk are highly baro-resistant
up to 300 MPa pressure treatments; however, at 500 MPa approximately 35% denaturation occurs.
It could be a strategy for preserving the colostrum immunoglobulins, which are heat labile (Felipe
et al., 1997). The pressure intensity of treatment is the major determinant of the structural stability
of Igs under hydrostatic pressure. Though immunoglobulins are baro-resistant, the critical pressure
reported to induce pressure-assisted inactivation is 275.9 MPa (Li et al., 2006), with 94% destruction
in activity at 689.7 MPa for 4 min. The rate of order of pressure-induced denaturation of immuno-
globulins is reported to be 1.5, but the effects of pressurization treatment (450–700 MPa at 20°C) in
different media viz. skim milk, rennet whey, and phosphate buffer were different, in the sense that
rate constant for pressure-induced denaturation was the least for skim milk whereas highest for whey,
indicating the protective effects of other milk constituents in immunoglobulins (Mazri et al., 2012).
5.4.3 Effect of high-pressure treatment on Milk Fat, Lactose, and Micronutrients
High-pressure treatment has been observed to induce crystallization in milk fat and an increase
in total solid content in cream. This is probably due to the fact that the phase transition from solid
to liquid of milk fat is shifted to higher values under pressure (15.5°C/100 MPa). In addition, lower
degree of milk fat crystallization at higher pressures (>350 MPa) may be due to a reduced molecular
mobility of this lipid under high-pressure conditions (Huppertz et al., 2002). Studies carried out by
Gervilla et al. (2001) on free fatty acids (FFA) content in ewe’s milk have showed that HP treatments
between 100 and 500 MPa at 4°C, 25°C, and 50°C did not increase FFA content. HP treatment at
higher processing temperature resulted in lower FFA values in the milk. The phenomenon is of
great interest to avoid production of off-flavours in milk and milk products, often encountered due
to lipolytic rancidity in milk. Hydrostatic pressure up to 500 MPa affects changes in size and distri-
bution of milk fat globules of ewe’s milk. HP treatments at 25°C and 50°C showed an increase in the
number of small globules in the range 1–2 μm, whereas at 4°C the tendency was reversed (Gervilla
et al., 2001). These changes on distribution of milk fat globules could be due to the phenomenon
of aggregation and disaggregation/disintegration and offer certain advantages for HP-treated milk.
HP treatment increases the stability of milk treated at 25°C and 50°C, whereas at 4°C it increases
the creaming-off, which could improve cream separation during butter manufacture.
During heating of milk lactose may isomerise to lactulose and then degrade to form acids and
other sugars. No changes in these compounds have been observed after pressurization in the range of
100–400 MPa for 10–60 min at 25°C, suggesting that no lactose isomerization or Maillard reaction
occurs in milk after pressure treatment (López-Fandiño et al., 1996). Contrary to thermal treatments,
where covalent as well as non-covalent bonds are affected, HP treatment at room and mild tempera-
tures only disrupts relatively weak chemical bonds (hydrogen bonds, hydrophobic bonds, ionic bonds).
Thus, small molecules such as vitamins, amino acids, simple sugars, and flavour compounds remain
unaffected by the HP treatment. HP treatment of milk at 400 MPa for 30 min at 25°C results in
non-significant loss of Vitamin B1 and B6 (Sierra et al., 2000). Studies with water-soluble vitamins in
fruits juices showed that pressure treatment had no significant effect on the levels of vitamin B and C.
5.5 hIGh-prESSUrE-MEDIatED EFFECt ON MILK ENZYMES
Inactivation of native enzymes by HP treatment has been attempted by several workers and quite
variable results obtained. The effects of HPP on inactivation of enzymes depend on the structure of
the enzyme and the processing conditions applied. Raw milk is reported to contain about 20 different
types of enzymes, which affect the quality of raw milk positively or adversely. Certain enzymes such
as lactoperoxidase have a major role in determining the microbial stability of raw milk. Likewise,
plasmin has a major role in cheese making. Alkaline phosphatase used as indicator for determining
the efficacy of pasteurization process appears quite pressure-resistant, with no inactivation in raw milk
after treatment at 400 MPa for 60 min at 20°C. The enzyme is completely inactivated at 800 MPa for
8 min. Similar observation was noticed for alkaline phosphatase in another study, where it remained
resistant to pressurization up to 400 MPa; but at higher pressure (600–800 MPa) and elevated temper-
ature inactivation enhanced. Several other enzymes like lactoperoxidase, phosphohexoseisomerase,
γ-glutamyltransferase, and plasmin have also been reported to resist HP treatment. Researchers have
indicated that lipoprotein lipase (LPL) is pressure stable. In fact, the activity of lipases in milk is
enhanced by HPP (Buffa et al., 2001). Combinations of higher pressure and temperature are necessary
to inactive lipases in milk. Pressure treatments at 400 MPa for 15 min at 40°C–60°C reduce the pro-
teolytic activity in milk. The reduced proteolytic activity or proteolysis is probably due to inactivation
of the plasmin system in milk at higher temperatures. There is also a synergistic effect of pressure and
temperature on the extent of proteolytic activity in milk. Proteolysis in milk during incubation at 37°C
is more extensive in milk treated at 300 MPa and room temperature (25°C) than at 400 MPa and 60°C.
The reasons for increased proteolytic activity are related to a combined effect towards greater disrup-
tion of casein micelles under pressure and little inactivation of plasmin at room temperature, resulting
in more surface area of exposure to proteolytic enzymes in pressure-treated milk (Huppertz et al.,
2002). Rademachera and Hinrichs (2006) studied the effect of high-pressure-induced inactivation
of indigenous milk lactoperoxidase (LP), phosphohexoseisomerase (PHI), and γ-glutamyltransferase
(GGT), and noticed that these are also stable at pressures up to 400 MPa at 20°C–25°C. With respect
to pressure stability the following ranking was observed: LP>GGT>PHI. García-Risco et al. (2000)
found that HP treatments at 400 MPa for 15 min at 40°C–60°C reduce the proteolytic activity, and at
25°C–60°C improve the organoleptic properties of milk, suggesting that these combined treatments
could be used to produce milk of good sensory properties with an increased shelf life.
5.6 hIGh-prESSUrE-rELatED ChaNGES ON

FUNCtIONaL CharaCtErIStICS OF MILK
Micelle disintegration induced by HP treatment also affects milk colour. A study was carried
out by Harte et al. (2003) to observe the series of changes during combined treatment of thermal and
HHP for yogurt manufacture and it was observed that milk subjected to HHP or thermal treatment
and then HHP lost its white colour and turned yellowish, which may be due to reduction in size of
casein micelles (Needs et al., 2000). On the other hand, milk when first subjected to HP and then
to thermal treatment regained its whitish colour, which is attributed to reversible nature of casein
micelles (or re-aggregation of disrupted micelles) towards HHP treatment when applied in the range
of 300–676 MPa followed by thermal treatment. HPP has been observed to affect the heat coagula-
tion time (HCT) of milk and pressure treatment above 200 MPa was found to decrease the HCT.
The effect on HCT could be attributed to HP-induced changes in casein and milk minerals.
Likewise, rennet coagulation time (RCT) of milk has also been observed to be influenced
by HPP; however, HP-induced changes in the rennet coagulation properties of milk appear quite
variable because of two opposite phenomena occurring in milk proteins on pressure treatment.
HP-induced disruption of casein micelles and HP-induced dissociation of micellar k-casein appear
to greatly reduce RCT and increase the rate of gelation and the strength of the gel. HP-induced asso-
ciation of denatured β-lactoglobulin with casein micelles hinders the rennet coagulation of milk,
resulting in increases (or smaller decreases) in RCT, and milder increases in gel strength and the rate
of gelation. The relative extent to which both these phenomena occur determines the outcome of a
treatment on the rennet coagulation properties of milk. However, milk pH and the concentration of
ionic calcium are two important factors affecting rennet coagulation (Zobrist et al., 2005), which
needs to be investigated in details. With the application of HP treatment as described, the size
and number of casein micelles increases as spherical particles change to form chains or clusters
of sub-micelles, thus reducing RCT. In general, rennet coagulation properties of milk subjected
to pressures of 100–500 MPa for 30 min are enhanced. Garcia-Risco et al. (2000) found that HP
treatments at 400 MPa for 15 min at 40°C–60°C reduced the proteolytic activity, and at 25°C–60°C
improved the organoleptic properties of milk, suggesting that these combined treatments could be
used to produce milk of good sensory properties with an increased shelf life.
Liu et al. (2005b) studied the effect of HHP treatments on hydrophobicity of whey protein concen-
trate (WPC) and observed that treatment of WPC yielded increase in the number of binding sites and
led to certain modifications of proteins and showed promising results for improving functional proper-
ties of foods. Similar observations for improved hardness, surface hydrophobicity, solubility, gelation,
and emulsifying properties were recorded by Lee and Kaletunc (2010) in whey protein functionality.
5.7 prOSpECtIVE appLICatIONS OF hIGh-prESSUrE

prOCESSING IN DaIrY INDUStrY
In dairy products, high pressure (up to 400 MPa) has been used to process milk successfully,
which has resulted in significant effect on milk molecules that could also influence the characteris-
tics of milk products like yogurt, cheese, cream, etc. (Chawla et al., 2011). The summarized appli-
cation of HP technology and related effect on product quality is depicted in Table 5.2. Some of the
prospective applications have been discussed below.
5.7.1 hhp Induced Effect on Cheese Making Characteristics
Milk pasteurization destroys pathogenic and almost all spoilage microorganisms and it is the
most important heat treatment applied to cheese milk to provide acceptable safety and quality.
However, milk pasteurization is known for its adverse effects with respect to many sensory char-
acteristics of cheese, leading to alterations in texture and often delayed maturation (Grappin and
Beuvier, 1997). HP technology can be used to increase the microbiological safety and quality of milk
to produce high quality cheeses. As has been mentioned previously, HP processing of milk at room
temperature causes several protein modifications, such as whey protein denaturation and micelle
fragmentation, and alters mineral equilibrium. It has been observed that denaturation of whey pro-
teins due to applied pressure results in interaction between denatured whey protein and casein, which
in turn increases the retention of former within casein matrix in cheese. Thus, these changes result in
modifying the technological aptitude of milk to make cheese, improving the rennet coagulation and
yield properties of cheese milk (San Martin-Gonzalez et al., 2004). Interest in applications of HP in
cheese processing has focused on areas such as inactivation or reduction of pathogenic and spoilage
microorganisms, acceleration of ripening, and increasing cheese yield. Various studies have evalu-
ated the potential of HP treatment of milk to improve the texture of low-fat cheese (Molina et al.,
2000) and treatment of cheese to reduce moisture variability within and between individual blocks
(Torres-Mora et al., 1996), and to improve rheological (Messens et al., 2000; Saldo et al., 2001) and
physiochemical characteristics (Capellas et al., 2001). Johnston and Darcy (2000) reported that HP
treatment of 1-day-old Mozzarella at 200 MPa for 60 min resulted in a reduction in hardness and an
increase in flow properties of the heated cheese. O’Reilly et al. (2002) attributed the positive influ-
ence of HP treatment on the cooking properties of Mozzarella to an increase in protein hydration and
swelling of the para-casein matrix of the cheese. Sheehan et al. (2005) reported that HP treatment
at 400 MPa for 5 min at 21°C did not significantly affect composition, pH, proteolysis, rheology, or
cooking properties of reduced-fat Mozzarella cheese over a 35-day storage period.
table 5.2 application of hhp in Dairy processing
product Key Feature references

Cheese Accelerated ripening of goat milk cheese at saldo et al. (2003)
400 MPa for 5 min in as compared to
conventional ripening
in Hispanico cheese manufactured using a Avila et al. (2006)
mixture of cows’ and ewes’ milk, HP treatment at
400 MPa for 5 min accelerated the proteolysis
without influencing sensory quality
HHP treatment at 300 MPa improved the ripening Juan et al. (2008)
of ewes’ milk cheese at all stage of ripening
enhancement of proteolysis was a function of o’reilly et al. (2000)
combination of the temperature and pressure
used in the treatment
Accelerated ripening of Cheddar cheese within yokoyama (1992)
3 days, comparable to 6-month of conventional
ripening, when treated at 50 MPa
shorter rennet coagulation at the pressure up to Johnston et al. (2002b)
250 MPa
incorporation of whey proteins upon HHP Lopez-Fandino et al. (1996)
treatment increases cheese yield up to 20%
elimination of non-starter microorganism during Gallot-Lavalle (1998)
ripening of cheese
elimination of bacteriophage completely at Capra et al. (2009)
100 MPa
Yogurt Faster acidification of HHP-treated milk as Huppertz et al. (2004b)
compared to control
Firm gel and resistant to syneresis Johnston et al. (2002a)
Prevent post-acidification and maintain viability of tanaka and Hatanaka (1992)
counts in yogurt
probiotic dairy products HHP-treated milk suitable for probiotic dairy food shah (2000)
manufacturing due to improved proteolysis by
poor proteolytic probiotic strains
Ice-cream Low-fat ice-cream with similar textural properties Lim et al. (2008)
to full-fat samples,
Partially denatured WPC solutions at 300 MPa for
15 min (25°C) and incorporated at 0.82% ts and
0.3% protein in low-fat ice-cream
no need of emulsifier and stabilizer Keenan et al. (1998)
slower melting rate and improved sensory
properties compared to the control
Microbiological quality of cheeses from HP-treated milk (500 MPa for 15 min at 20°C) was com-
parable to pasteurized milk (72°C for 15 s) cheeses (Buffa et al., 2001). However, the application of
HP technology to cheese milk causes differences in cheese composition and ripening in comparison
to pasteurized milk cheese. The HP-treated milk cheeses have higher moisture, salt, and total free
amino acids contents than raw or pasteurized milk cheeses. On the other hand, cheeses made from
HP-treated milk showed a similar level of lipolysis to cheeses made from raw milk, whereas the level
of lipolysis in cheese made from pasteurized milk was lower and this behaviour was explained by heat-
sensitive but partial pressure-resistant characteristics of the indigenous milk lipase. Also, pressure-
treated cheese showed more viscoelastic texture and had less resistance to flow (Messens et al., 2000).
Being that cheese ripening is a quite expensive process, acceleration of ripening is highly
desirable. Most of the work in this field has been done by elevation of ripening temperature, addi-
tion of cheese slurries or exogenous enzymes, or by the use of adjunct starters, either as such or
in modified form. Experimental Cheddar cheese samples were exposed to pressure from 0.1 to
300 MPa for 3 days at 25°C after cheese making. Best results were obtained at 50 MPa, where
cheese with free amino acid and taste comparable to that of a 6-month-old commercial cheese was
obtained. In particular, the kind of starter bacteria added to the cheese milk was highly proteolytic
and at least 10-fold higher than conventional inoculation rates. In certain cheese varieties such as
Mozzarella and Gouda, pressurization increases rate of proteolysis on exposure to pressure treat-
ment of 400–600 MPa for 5–15 min. There is substantial scope for improving the quality of cheeses
made from Buffalo milk.
5.7.2 Yoghurt and Ice-Cream
Yoghurt, a very popular dairy product, suffers from the common defect of syneresis and low
viscosity or rigidity. Yoghurt quality can be improved in terms of its preservation and improved
rheological properties by pressurization treatment. Skim milk treated with combined treatments
of HHP (400–500 MPa) and thermal treatment (85°C for 30 min) showed increased yield stress,
resistance to normal penetration, elastic modulus, and reduced syneresis (Harte et al., 2003).
High-pressure-treated milk has been successfully used to manufacture low-fat set-type yoghurt
with a creamy, thick consistency and there was no requirement of any hydrocolloids to overcome
the textural defects (Moorman et al., 1996). The application of high pressure to full-fat yoghurts
to check post-acidification after packaging has also been achieved (Tanaka and Hatanaka, 1992).
They observed that treatments at 200–300 MPa for 10 min at room temperature prevented acidi-
fication and maintained the initial number of viable lactic acid bacteria and the yoghurt texture.
Reps et al. (1999) investigated the effect of pressurization on inactivation of microflora present in
yogurt and found that HP treatment of 400 MPa completely inactivates Lactobacillus bulgaricus,
but Streptococcus thermophilus has been found to be more resistant towards pressure and its resis-
tance varies from strain to strain, in the range of 35.3%–99.9%, which will be beneficial in checking
post-acidification in yoghurt, a major detriment for longer shelf life of that food.
HPP treatment induced fat crystallization, shortened the time required to achieve a desirable
solid fat content, thereby reducing the ageing time of ice-cream, and also enhanced the physical
ripening of cream for making butter (Buchheim and Frede, 1996).
5.7.3 prospective applications in Functional Dairy Foods
Colostrum is the mammary secretion produced during the first 72 h after parturition that pro-
vides nourishment for the new-born. It contains numerous immune system and growth factors
as well as essential nutrients, trypsin, and protease inhibitors that protect it from destruction in
the gastrointestinal tract. The most important components of colostrum can basically be divided
into two major categories: immune factors and growth factors. Colostrum is a complex fluid,
obtained after calving of animals, which possesses higher levels of nutrients and bioactive compo-
nents, e.g., immunoglobulins (Igs), especially IgG1; growth factors, especially insulin-like growth
factors-1 (IGF-1); transforming growth factor beta-2 (TGF-β2) and growth hormone (GH); as well
as lactoferrin, lysozyme, and lactoperoxidase. Colostral antibodies provide passive immunity to the
new-born calf and growth factors control some fundamental life processes such as cell division, cell
differentiation, or apoptosis, and stimulate the growth and development of the gastrointestinal tract
of new-born animals. The major Ig present in ruminant milk is IgG, with IgG1 representing more
than 90%, but also IgA and IgM are found. The level of these bioactive substances in colostrums and
colostrum fractions after processing operation including membrane processing, heat treatment, and
freeze-drying decreased gradually. Borad and Singh (2018) compiled the processing-related aspects
of colostrum and concluded that HP processing of colostrum could be an effective way of retaining
these bioactive components in colostrum without compromising with the safety and shelf-life.
Microfiltration followed by HPP (400 and 500 MPa for 10 min) to skim colostrum yielded practical
sterility without detection of any microbial load (Gosch et al., 2014) and hence would be recom-
mended to process and preserve heat-sensitive biological fluids as HPP offered the least detrimen-
tal effect on immunoglobulins as compared to thermal processing required to achieve equivalent
microbial reduction. This could be due to ability of immunoglobulins to form pressure-induced
compressed gel, identical to thermal gel, with retained bioactivity due to retained native structure
(George et al., 2013). Li et al. (2006) reported 20% more retention in immunoglobulin G activity at
HPP as compared to thermal processing (78°C/120 s) at 5-log reduction in microbial load. The batch
pasteurization reduced immunoglobulin A to 28% of original load, as compared to total retention at
HPP at 400 MPa, 87.9% retention at 500 MPa, and 69.3% retention at 600 MPa, respectively.
Colostrum has antioxidant and anti-inflammatory properties and is a good source of many vita-
mins, minerals, enzymes, and amino acids. Heat treatment and freeze drying are the two most
common processing technologies which have been applied so far for preserving the colostrum, but
these cause substantial losses of immunoglobulins, growth factors, and therapeutic minor whey
proteins. There is increasing demand for colostrum powder because of its health-promoting proper-
ties for infants, women, and persons suffering with immune disorders including HIV and cancer.
HHP technology has been reported to have minor effect on bioactivity of colostrum preparations,
but there is need for systematic and detailed investigations.
Probiotic foods are one of the fastest growing sectors of functional foods. One of the major
obstacles in efficacy of probiotic dairy products is the loss of viability of microorganisms dur-
ing processing, storage, and consumption. Irrespective of the preservation method applied, pro-
biotic cultures are exposed to unfavourable environmental conditions, due to increased solute
concentration, intracellular ice formation in case of freezing and freeze drying, exposure to high
temperatures during spray drying, and dehydration. Penna et al. (2006) investigated the effect of
using HP (676 MPa for 5 min) and heat treatment (85°C for 30 min) alone on milk to be used for the
production of probiotic yogurts with two different starters and Bifidobacterialongum. Application
of HP led to compact yogurt gel with increasingly larger casein micelle clusters interspaced by void
spaces, and exhibited a high degree of cross-linking. On the other hand, in yogurt made with high
heat-treated milk, the micelles were less interconnected and exhibited irregular shapes with large
pores. Irrespective of the strain used, yogurts made from HP milk and those with combination of
thermal and HP treatment had lower syneresis, compared to yogurts made from thermally treated
milk. Probiotic yogurts made from HP milk yielded gels with higher consistency index than gels
obtained from thermally treated milk. Moreover, gel firmness of the yogurt was depended on the
starter culture employed (Penna et al., 2006, 2007). The treatments did not affect the products’
physicochemical properties or the starter and probiotic bacteria counts.
HP and PEF, particularly, might be highly useful techniques employed in the preservation of
ingredients used in processing liquid probiotic foods (Pereira and Vicente, 2010). In HPP, the food
(solid or liquid) is subjected to pressures above 100 MPa up to 900 MPa, with pressures used in
commercial systems between 400 and 700 MPa. The probiotic microorganism should be technologi-
cally suitable for incorporation in the food product retaining their viability and efficacy in the food
product. There is great scope for utilizing HP processing for the development of probiotic dairy
foods with higher viable counts.
5.8 CONCLUSION
HPP products are becoming the choice of a new consumer in terms of health and safety aspects.
Being one of the emerging technologies in developing countries, high-pressure technology offers the
technologists an opportunity to develop novel products with enhanced shelf life and food safety, bet-
ter sensory and nutritional quality of food, and feasibility to process a wide range of products. It also
offers food processors a scope to manufacture minimally processed shelf-stable products. However,
the potential of high-pressure treatment for shelf-life extension without affecting the nutritional value
and modification of functionalities of macromolecules for exploitation of improving the quality char-
acteristics of resultant products needs to be investigated. HPP offers considerable scope for delivering
high-value dairy products such as colostrum, probiotic dairy products, infant foods, and even shelf-life
enhancement of human milk without affecting its nutritional and therapeutic virtues.
rEFErENCES
Aoki, T., Yamada, N., and Kako, Y. (1990). Relationship between colloidal calcium phosphate cross-linkage
and release of β-casein from bovine casein micelles on cooling. Agr. Biol. Chem. Tokyo. 54: 2287–2292.
Arias, M., Lopez-Fandino, R., and Olano, A. (2000). Influence of pH on the effects of high pressure on milk
proteins. Milchwissenschaft. 55: 191–194.
Avila, M., Garde, S., Gaya, P., Medina, M., and Nunez, M. (2006). Effect of high-pressure treatment and a
bacteriocin-producing lactic culture on the proteolysis, texture, and taste of Hispanico cheese. J. Dairy
Sci. 89: 2882–2893.
Balasubramanian, S., and Balasubramanian, V.M. (2003). Compression heating influence of pressure trans-
mitting fluids on bacteria inactivation during high pressure processing. Food Res. Int. 36(7): 661–668.
Benito, A., Ventoura, G., Casadei, M., Robinson, T., and Mackey, B. (1999). Variation in resistance of natu-
ral isolates of Escherichia coli O157 to high hydrostatic pressure, mild heat, and other stresses. Appl.
Environ. Microbiol. 65: 1564–1569.
Bonomi, F., Fiocchi, A., Frokiaer, H., Gaiaschi, A., Iametti, S., Poiesi, C., Rasmussen, P., Restain, P., and
Rovere, P. (2003). Reduction of immunoreactivity of bovine β-lactoglobulin upon combined physical
and proteolytic treatment. J. Dairy Res. 70: 51–59.
Borad, S.G., and Singh, A.K. (2018). Colostrum immunoglobulins: Processing, preservation and application
aspects. Int. Dairy J. 85: 201–210
Borad, S.G., Kumar, A., and Singh, A.K. (2017). Effect of processing on nutritive values of milk protein. Crit.
Rev. Food Sci. Nutri. 57(17): 3690–3702.
Buchheim, W., and Abou El-Nour, A.M. (1992). Induction of milk fat crystallization in the emulsified state by
high hydrostatic pressure. Fat Sci. Technol. 94: 369–373.
Buchheim, W., and Frede. E. (1996). Use of high-pressure treatment to influence the crystallisation of emulsi-
fied fats. DMZ Lebensmittel Industrie and Milchwirtschaft. 117(5): 228–237.
Buchheim, W., Schrader, K., Morr, C.V., Frede, E., and Schutt, M. (1996). Effects of high pressure on the
protein, lipid and mineral phase of milk. In: Proceedings of the IDF Symposium Heat Treatments and
Alternative Methods, pp. 202–213. Special Issue No 9602. Brussels, Belgium: International Dairy
Federation.
Buffa, M., Guamis, B., Pavia, M., and Trujillo, A.J. (2001). Lipolysis in cheese made from raw, pasteurized or
high-pressure treated goat’s milk. Int. Dairy J. 11: 175–179.
Capellas, M., Mor-Mur, M., Sendra E., and Guamis, B. (2001). Effect of high-pressure processing on physico-
chemical characteristics of fresh goat’s milk cheese (Mato). Int. Dairy J. 11: 165–173.
Capra, M.J., Patrignani, F., Quiberoni, A.L., Reinheimer, J.A., Lanciotti, R., and Guerzoni, M.E. (2009).
Effect of high pressure homogenization on lactic acid bacteria phages and probiotic bacteria phages. Int.
Dairy J. 19(5): 336–341.
Chawla, R., Patil, G.R., and Singh, A.K. (2011). High hydrostatic technology in dairy processing. J. Food Sci.
Technol. 48(3): 260–268.
Cheftel, J.C. (1995). Review: High-pressure, microbial inactivation and food preservation. Food Sci. Technol.
Int. 1: 75–90.
Chicon, R., Lopez-Fandino, R., Alonso, E., and Belloque, J. (2008). Proteolytic pattern, antigenicity, and
serum immunoglobulin E binding of β-lactogobulin hydrolysates obtained by pepsin and high pressure
treatments. J. Dairy Sci. 91: 928–938.
Dufour, E., Herve, G., and Haertle, T. (1995). Hydrolysis of β-lactoglobulin by thermolysin and pepsin under
high hydrostatic pressure. Biopolymers. 35: 475–483.
Felipe, X., Capellas, M., and Law, A.R. (1997). Comparison of the effects of high-pressure treatments and heat
pasteurisation on the whey proteins in goat’s milk. J. Agric. Food Chem. 45(3): 627–631.
Fonberg-Broczek, M., Arabas, J., Kostrzewa, E., Reps, A., Szczawinski, J., Szczepek, J., Windyga, B., and
Porowski, S. (1999). High pressure treatment of fruit, meat, and cheese products: equipment, meth-
ods and results. In: Processing Foods, Quality Optimisation and Process Assessment, pp. 281–300.
Oliveira, F.A.R., and Oliveira, J.C., Eds., CRC Press LLC, Boca Raton, FL.
Frede, E., and Buchheim, W. (2000). The influence of high pressure upon the phase transition behaviour of
milk-fat and milk-fat fractions. Milchwissenschaft. 55: 683–686.
Gallot-Lavallee, T. (1998). Efficiency of high pressure treatment for the destruction of Listeria monocytogenes
in goat cheese from raw milk. Sci. Aliments. 18(6): 647–655.
Garcia-Risco, M.R., Recio, I., Molina, E., and Lopez-Fandino, R. (2003). Plasmin activity in pressurized
milk. J. Dairy Sci. 86: 728–734.
Garcia-Risco, M.R., Olano, A., Ramos, M., and Lopez-Fandino, R. (2000). Micelar changes induced by high
pressure. Influence in the proteolytic activity and organoleptic properties of milk. J. Dairy Sci. 83(10):
2184–2189.
Gaucheron, F., Famelart, M.H., Mariette, F., Raulot, K., Michel, F., and Le Graet, Y. (1997). Combined effects
of temperature and high pressure treatments on physicochemical characteristics of skim milk. Food
Chem. 59: 439–447.
George, P., Kasapis, S., Bannikova, A., Mantri, N., Palmer, M., Meurer, B., and Lundin, L. (2013). Effect of
high hydrostatic pressure on the structural properties and bioactivity of immunoglobulins extracted
from whey protein. Food Hydrocolloids. 32: 286–293.
Gervilla, R., Ferragut, V., and Guamis, B. (2000). High pressure inactivation of microorganisms inoculated
into ovine milk of different fat contents. J. Dairy Sci. 83(4): 674–682.
Gervilla, R., Ferragut, V., and Guamis, B. (2001) High hydrostatic pressure effects on colour and milk-fat
globule of ewes’ milk. J. Food Sci. 66: 880–885.
Gosch, T., Apprich, S., Kneifel, W., and Novalin, S. (2014). A combination of microfiltration and high pressure
treatment for the elimination of bacteria in bovine colostrum. Int. Dairy J. 34(1): 41–46.
Gould, G.W. (2000). Preservation: Past, present and future. Br. Med. Bull. 56: 84–96.
Gould, G.W., and Sale, A.J.H. (1970). Initiation of germination of bacterial spores by hydrostatic pressure.
J. Gen. Microbiol. 60: 335–346.
Grappin, R., and Beuvier, E. (1997). Possible implications of milk pasteurisation on the manufacture and
sensory quality of ripened cheese. Int. Dairy J. 7(12): 751–761.
Harte, F.M., Gurram, S.R., Luedecke, L., Swanson, B.G., and Barbosa-Canovas, G.V. (2007). Effect of high
hydrostatic pressure and whey proteins on the disruption of casein micelle isolates. J. Dairy Res. 74:
452–458.
Harte, F., Luedecke, L., Swanson, B., and Barbosa-Canovas, G.V. (2003). Low-fat set yogurt made from
milk subjected to combinations of high hydrostatic pressure and thermal processing. J. Dairy Sci. 86:
1074–1082.
Hayman, M., Kouassi, G.K., Anantheswaran, C.R., Floros, D.J., and Stephen, J.K. (2008). Effect of water
activity on inactivation of Listeria monocytogenes and lactate dehydrogenase during high pressure pro-
cessing. Int. J. Food Microbiol. 124: 21–26.
Hinrichs, J., and Rademacher, B. (2004). Kinetics of combined thermal and pressure induced whey protein
denaturation in bovine skim milk. Int. Dairy J. 14: 315–323.
Hinrichs, J., Rademacher, B., and Kessler, H.G. (1996a). Food processing of milk products with ultrahigh
pressure. In: Heat Treatments and Alternative Methods, pp. 185–201. Special Issue No. 9602. Brussels,
Belgium: International Dairy Federation.
Hinrichs, J., Rademacher, B., and Kessler, H.G. (1996b). Reaction kinetics of pressure induced denaturation of
whey proteins. Milchwissenschaft. 51: 504–509.
Hite, B.H. (1899). The effect of pressure in the preservation of milk. West Virginia University Agricultural and
Experimental Station Bulletin. 54: 15–35.
Holt, C., Timmins, T.A., Errington, N., and Leaver, J. (1998). A core-shell model of calcium phosphate nanoclus-
ters derived from sedimentation equilibrium and small angle X-ray and neutron scattering measurements.
Eur. J. Biochem. 252: 73–78.
Huppertz, T., Fox, P.F., and Kelly, K.L. (2004a). High pressure treatment of bovine milk: Effects on casein
micelles and whey proteins. J. Dairy Res. 71: 97–106.
Huppertz, T., Fox, P.F., and Kelly, A.L. (2004b). Influence of high pressure treatment on the acidification of
bovine milk by lactic acid bacteria. Milchwissenschaft. 59: 246–249.
Huppertz, T., Fox, P.F., and Kelly, K.L. (2004c). High pressure-induced denaturation of α-lactalbumin and
β-lactoglobulin in bovine milk and whey: A possible mechanism. J. Dairy Res. 71: 489–495.
Huppertz, T., Kelly, A.L., de Kruif, K.G., and Fox, P.F. (2006a). High pressure induced changes in bovine milk
proteins: A review. Biochim. Biophys. Acta. 1764: 593–598.
Huppertz, T., Kelly, A.L., and Fox, P.F. (2002). Effect of high pressure on constituents and properties of milk.
Int. Dairy J. 12: 561–572.
Huppertz, T., Kelly, A.L., and Fox, P.F. (2006b). High pressure induced changes in ovine milk: Effects on
casein micelles and whey proteins. Milchwissenschaft. 61: 394–397.
Jaenicke, R. (1981). Enzymes under extreme conditions. Ann. Rev. Biophy. Bioeng. 10(1): 1–67.
Johnston, D.E., and Darcy P.C. (2000). The effects of high pressure treatment on immature Mozzarella cheese.
Milchwissenschaft. 55: 617–620.
Johnston, D.E., Murphy, R.J., Rutherford, J.A., and McCreedy, R.W. (2002a). Acidification of high pressure
treated milk: The role of whey protein denaturation. Milchwissenschaft. 57: 605–608.
Johnston, D.E., Rutherford, J.A., and McCreedy, R.W. (2002b). Ethanol stability and chymosin-induced coag-
ulation behaviour of high pressure treated milk. Milchwissenschaft. 57: 363–366.
Juan, B., Ferragut, V., Guamis, B., and Trujillo, A.J. (2008). The effect of high-pressure treatment at 300 MPa
on ripening of ewes’ milk cheese. Int. Dairy J. 18(2): 129–138.
Keenan, R.D., Wix, L., and Young, D. (1998). Method for the Preparation of a Foodstuff. Patent WO/98/18350.
Kleber, N., Maier, S., and Hinrichs, J. (2007). Antigenic response of bovine β-lactoglobulin influenced by
ultrahigh pressure treatment and temperature. Innov. Food Sci. Emerg. Technol. 8: 39–45.
Knorr, D. (1995). Hydrostatic pressure treatment of food: Microbiology. In: New methods of food preserva-
tion, pp 159–175. Gould, G.W. Eds., Blackie Academic and Professional, London.
Law, A.J.R., Leaver, J., Felipe X., Ferragut, V., Pla, R., and Guamis, B. (1998). Comparison of the effects of high
pressure and thermal treatments on the casein micelles in goat’s milk. J. Agri. Food. Chem. 46(7): 2523–2530.
Ledenbach, L.H., and Marshall, R.T. (2009). Microbiological Spoilage of Dairy Products. In: Compendium of
the Microbiological Spoilage of Foods and Beverages, pp. 41–67. Sperber W., Doyle M., Eds., Springer,
New York, NY.
Lee, L., and Kaletunc, G. (2010). Inactivation of Salmonella Enteritidis strains by combination of high hydro-
static pressure and nisin. Int. J. Food Microbiol. 140(1): 49–56.
Li, S.Q., Zhang, H.Q., Balasubramaniam, V.M., Lee, Y.Z., Bomser, J.A., Schwartz, S.J., and Dunne, C.P.
(2006). Comparison of effects of high-pressure processing and heat treatment on immunoactivity of
bovine milk immunoglobulin G in enriched soymilk under equivalent microbial inactivation levels.
J. Agri. Food Chem. 54(3): 739–746.
Lim, S.Y., Swanson, B.G., Ross, C.F., and Clark, S. (2008). High hydrostatic pressure modification of whey
protein concentrate for improved body and texture of low fat ice cream. J. Dairy Sci. 91(4): 1308–1316.
Liu, X., Powers, J.R., Swanson, B.G., Hill, H.H., and Clark, S. (2005a). High hydrostatic pressure affects
flavor-binding properties of whey protein concentrate. J. Food Sci. 70: C581–C584.
Liu, X., Powers, J.R., Swanson, B.G., Hill, H.H., and Clark, S. (2005b). Modification of whey protein con-
centrate hydrophobicity by high hydrostatic pressure. Innov. Food Sci. Emerg. Technol. 6(3): 310–317.
López-Fandiño, R., Carrascosa, A.V., and Olano, A. (1996). The effects of high pressure on whey protein
denaturation and cheese-making properties of raw milk. J. Dairy Sci. 79: 929–936.
López-Fandino, R., De la Fuente, M.A., Ramos, M., and Olano, A. (1998), Distribution of Minerals and
Proteins Between the Soluble and Colloidal Phases of Pressurized Milks from Different Species.
J. Dairy Res. 65, 69–78.
Mazri, C., Ramos, S.J., Sánchez, L., Calvo, M., and Pérez, M.D. (2012). Reaction kinetics of pressure-induced
denaturation of bovine immunoglobulin G. Int. Dairy J. 24: 8–12.
Messens, W., Foubert, I., Dewettinck, K., and Huyghebaert, A. (2000). Proteolysis of a high-pressure-treated
smear-ripened cheese. Milchwissenschaft. 55(6): 328–332.
Messens, W., Van Camp, J., and Huyghebaert, A. (1997). The use of high pressure to modify the functionality
of food proteins. Trends Food Sci. Technol. 8: 107–112.
Molina, E., Alvarez, M., Ramos, M., Olano, A., and Lopez-Fandino, R. (2000). Use of high-pressure-treated
milk for the production of reduced-fat cheese. Int. Dairy J. 10: 467–475.
Moorman, J.E., Toledo, R.T., and Schmidt, K. (1996). High-pressure throttling (HPT) reduces population,
improves yogurt consistency and modifies rheological properties of ultrafiltered milk. IFT Annual
Meeting 1996: 49 p.
Mussa, D. M., and Ramaswamy, H. S. (1997). Ultra high pressure pasteurization of milk: Kinetics of microbial
destruction and changes in physico-chemical characteristics. LWT-Food Sci. Technol. 30(6): 551-557.
Needs, E.C., Stenning, R.A., Gill, A.L., Ferragut, V., and Rich, G.T. (2000). High pressure treatment of milk:
Effects on casein micelle structure and on enzymic coagulation. J. Dairy Res. 67(1): 31–42.
O’Reilly, C.E., Kelly, A.L., Murphy, P.M., and Beresford, T.P. (2001). High pressure treatment: Applications
in cheese manufacture and ripening. Trends Food Sci. Technol. 12(2): 51−59.
O’Reilly, C.E., Murphy, P.M., Kelly, A.L., Guinee, T.P., Auty, M.A.E., and Beresford, T.P. (2002). The effect of
high pressure treatment on the functional and rheological properties of Mozzarella cheese. Innov. Food
Sci. Emerg. Technol. 3: 3–9.
O’Reilly, C.E., O’Connor, P.M., Murphy, P.M., Kelly, A.L., and Beresford, T.P. (2000). The effect of exposure
to pressure of 50 MPa on Cheddar cheese ripening. Innov. Food Sci. Emerg. Technol. 1(2): 109–117.
Penas, E., Prestamo, G., Baeza, M.L., and Martınez-Molero, M.I. (2006). Effects of combined high pressure and
enzymatic treatments on the hydrolysis and immunoreactivity of dairy whey proteins. Int. Dairy J. 16: 831–839.
Penna, A.L.B., Subbarao, G., and Barbosa-Canovas, G.V. (2006). Effect of high hydrostatic pressure process-
ing on rheological and textural properties of probiotic low-fat yoghurt fermented by different starter
cultures. J. Food Process Eng. 29: 447–461.
Penna, A.L.B., Subbarao, G., and Barbosa-Canovas, G.V. (2007). High hydrostatic pressure processing on
microstructure of probiotic low-fat yoghurt. Food Res. Int. 40: 510–519.
Pereira, R.N., and Vicente, A.A. (2010). Environmental impact of novel thermal and non-thermal technologies
in food processing. Food Res Int. 43(7): 1936–1943.
Rademacher, B., and Hinrichs, J. (2006). Effects of high pressure treatment on indigenous enzymes in bovine
milk: Reaction kinetics, inactivation and potential application. Int. Dairy J. 16(6): 655–661.
Rademacher, B., and Kessler, H.G. (1997). High pressure inactivation of microorganisms and enzymes in milk
and milk products. In: High pressure bioscience and biotechnology, pp 291–293. Heremans, K. Eds.,
Leuven University Press, Leuven, Belgium.
Reps, A., Warminska-Radyko, I., and Dajnowiec, F. (1999). Effect of high pressure on yoghurt. In: Advances in
High Pressure Bioscience and Biotechnology, pp. 453–456. Ludwig, H., Ed., Springer, Berlin, Germany.
Saldo, J., Fernandez, A., Sendra, E., Butz, P., Tauscher, B., and Guamis, B. (2003). High pressure treatment
decelerates the lipolysis in a caprine cheese. Food Res. Int. 36: 1061–1068.
Saldo, J., Sendra, E., and Guamis, B. (2001). Hard cheese structure after a high hydrostatic pressure treatment
at 50 MPa for 72 h applied to cheese after brining. Le Lait. 81(5): 625–635.
San Martin-Gonzalez, M.F., Welti-Chanes, J.S., and Barbosa-Canovas, G.V. (2004). Cheese manufacturing
assisted by ultra-high pressure. IFT Meeting, July, pp. 12–16, Las Vegas, NV.
Schrader, K., Buchheim, W., and Morr, C.V. (1997). High pressure effects on the colloidal calcium phosphate
and the structural integrity of micellar casein in milk. Part 1. High pressure dissolution of colloidal
calcium phosphate in heated milk systems. Nahrung, 41: 133–138.
Shah, N.P. (2000). Probiotic bacteria: Selective enumeration and survival in dairy foods. J. Dairy Sci. 83(4):
894–907.
Sheehan, J.J., Huppertz, T., Hayes, M.G., Kelly, A.L., Beresford, T.P., and Guinee, T.P. (2005). High pressure
treatment of reduced-fat Mozzarella cheese: Effects on functional and rheological properties. Innov.
Food Sci. Emerg. Technol. 6: 73–81.
Sierra, I., Vidal, V., and Lopez, F. (2000). Effect of high pressure on the Vitamin B1 and B6 content in milk.
Milchwissenschaft. 55(7): 365–367.
Smelt, J.M. (1998). Recent advances in the microbiology of high pressure processing. Trends Food Sci.
Technol. 9(4): 152–158.
Stapelfeldt, H., Petersen, P.H., Kristiansen, K.R., Qvist, K.B., and Skibsted, L.H. (1996). Effect of high hydro-
static pressure on the enzymatic hydrolysis of β-lactoglobulin B by trypsin, thermolysin and pepsin.
J. Dairy Res. 63: 111–118.
Tanaka, T., and Hatanaka, K. (1992). Application of hydrostatic pressure to yogurt to prevent its
after-acidification. J. Jpn. Soc. Food Sci. Technol. 39(2): 173–177.
Tauscher, B. (1995). Pasteurization of food by hydrostatic high pressure: Chemical aspects. Z. Lebensmittel-
Untersuchung und -Forschung, 200: 3–13.
Toepfl, S., Mathys, A., Heinz, V., and Knorr, D. (2006). Potential of high hydrostatic pressure and pulsed
electric fields for energy efficient and environmentally friendly food processing. Food Rev. Int. 22(4):
405–423.
Torres-Mora, M.A., Soeldner, A., Ting, E.Y., Hawes, A.C.O., Alemán, G.D., Bakski, G. S., … and Torres, J. A.
(1996). Early microstructure changes in Cheddar cheese and effects of high pressure curd processing.
IFT Annu. Mtg. Tech. Program Abstr. 9.
Trujillo, A.T., Capellas, M., Saldo, J., Gervilla, R., and Guamis, B. (2002). Applications of high hydrostatic
pressure on milk and dairy products: A review. Innov. Food Sci. Emerg. Technol. 3: 295–307.
Vercammen, A., Vivijs, B., Lurquin, I., and Michiels, C.W. (2012). Germination and inactivation of Bacillus
coagulans and Alicyclobacillus acidoterrestris spores by high hydrostatic pressure treatment in buffer
and tomato sauce. Int. J. Food Microbiol. 152(3): 162–167.
Vachon, J.F., Kheadr, E.E., Giasson, J., Paquin, P., and Fliss, I. (2002). Inactivation of foodborne pathogens in
milk using dynamic high pressure. J. Food Protection. 65(2): 345–352.
Yokoyama, H., Sawamura, N., and Motobayashi, N. (1992). Method for accelerating cheese ripening. European
Patent EP 4 698 570.
Zeece, M., Huppertz, T., and Kelly, A. (2008). Effect of high-pressure treatment on in-vitro digestibility of
β-Lactoglobulin. Innov. Food Sci. Emerg. Technol. 9: 62–69.
Zobrist, M.R., Huppertz, T., Uniacke, T., Fox, P.F., and Kelly, A.L. (2005). High-pressure-induced changes in
the rennet coagulation properties of bovine milk. Int. Dairy J. 15: 655–662.
ChaptEr 6
pulsed Electric Field processing

Principles and Engineering Aspects
R. Singh, B. P. Kaur, and S. Thangalakshmi
CONtENtS
6.1 Introduction ............................................................................................................................ 89

6.2 Principle of PEF......................................................................................................................90
6.3 Mechanism of Microbial Inactivation ....................................................................................90
6.4 PEF Equipment ....................................................................................................................... 91
6.4.1 Treatment Chamber ....................................................................................................92
6.4.2 Types of Pulses ...........................................................................................................92
6.4.3 Pulse Characteristics ..................................................................................................92
6.4.4 Pulse Generation Networks ........................................................................................94
6.4.4.1 Conventional Techniques .............................................................................94
6.4.4.2 Power Electronics Based Technique ............................................................ 95
6.5 Factors Contributing to the Efficacy of PEF Technology .......................................................96
6.5.1 Technological Factors .................................................................................................96
6.5.1.1 Electric Field Strength (E) ...........................................................................96
6.5.1.2 Duration of Treatment..................................................................................97
6.5.2 Biological Factors .......................................................................................................97
6.5.3 Media Factors .............................................................................................................97
6.6 Kinetic Models for PEF Processing........................................................................................ 98
6.7 Application on Food Quality .................................................................................................. 98
6.7.1 Milk and Milk Products ............................................................................................. 98
6.7.2 Fruits and Vegetable Products .................................................................................. 101
6.7.3 Other Food Products ................................................................................................. 102
6.8 Conclusions ........................................................................................................................... 103
References ...................................................................................................................................... 103
6.1 INtrODUCtION
When foods are treated by conventional, thermal methods of preservation, several undesir-
able changes occur, including loss of nutritive value, flavour, texture, etc. Nowadays consumers
demand processed foods with preserved sensory, physical, and chemical quality. As alternatives to
traditional thermal treatments, several non-thermal pasteurization methods have been developed in
order to obtain safe, nutritive, and “fresh-like” food. Several processing techniques like ultrasound
89
treatment, ultraviolet irradiation, gamma irradiation, treatment with non-conventional chemical

reagents, high-intensity magnetic field, high-pressure processing, use of membrane technology, and
application of high-voltage pulsed electric field (PEF) have been investigated to attain sufficient
microbial reduction while maintaining food quality.
PEF is a novel non-thermal processing technology which applies the concept of applying high-
intensity electrical pulses to food products in order to achieve microbial inactivation and further
stored under refrigeration with proper packing mechanism resulting in increase of shelf life of the
food product (Janositz et al., 2011). This technology aims to provide better quality food to consumer
by killing microorganisms and is considered to be superior to traditional thermal processing methods
in preserving the food quality attributes, viz. colour, flavour, texture, and nutritional value of food.
6.2 prINCIpLE OF pEF
PEF involves application of an external electrical field to liquid or semi-solid food placed
between two electrodes for a few microseconds; it induces structural changes and a rapid break-
down of the cell membrane. This phenomenon is called electroporation, and several applications of
high-intensity PEF have been studied in the last decades on foods (Toepfl et al., 2005).
The principle of PEF processing involves application of pulses of high electric fields to liquid or
semi-solid food placed between two electrodes for a few microseconds to milliseconds and intensity
in the order of 10–80 kV/cm with the aim of inactivating microorganism, altering enzymes, inten-
sifying some methods like dehydration, and drying or attaining some specific transformation in
food product. PEF induces structural changes and a rapid breakdown of the cell membrane, which
is known as electroporation, due to which there is increase in permeability and electrical conductiv-
ity of cellular material. The high-voltage pulses cause temporary destabilization and perforation of
lipid bilayer and proteins but as electric field is removed, the pores can reseal themselves if the pores
induced are small in comparison to membrane area. If strength of the electric field (E) and treatment
intensity increase by increasing the pulse width and/or number of pulses, then there is formation
of irreversible pores due to which the microorganism will not be able to repair itself and will start
leaking small compounds; eventually cell death occurs (Amiali et al., 2006, Ortega-Rivas, 2012).
The magnitude of the voltage required for cell breakdown depends upon cell type, size, diam-
eter, shape, growth phase of microorganism, and medium in which the organism is present. Usually,
the trans-membrane voltage um induced on the cell membrane due to an external electric field is
given as (Zimmermann, 1986; Ortega-Rivas, 2012)
u m = αd c E cos θ (6.1)
where α depends on cell shape (α = 1 for rectangular cell; α = 0.75 for spherical cell), dc is the
cell diameter, E is the electric field strength, and θ is the angle between a point on the membrane
surface and the direction of electric field. The average diameters of microorganism and biologi-
cal tissue cells vary from 10 nm to 1 μm and 10 μm to 1 mm, respectively (Aguilera et al., 2000;
Ortega-Rivas, 2012).
6.3 MEChaNISM OF MICrOBIaL INaCtIVatION
The microorganism’s inactivation by PEF is associated mainly to the breakdown of the cell
membrane and its electromechanical instability (Coster and Zimmermann, 1975; Jacob et al., 1981).
Formation of pores on cell membrane of microorganism by high-intensity PEF pulses has not been
PuLseD eLeCtriC FieLD ProCessinG 91
Figure 6.1 Dielectric breakdown mechanism of microbial inactivation. ec is the critical electric field (a) intact
cell membrane, (b) membrane compression, (c) pore formation with reversible breakdown,
(d) irreversible breakdown.
Figure 6.2 electroporation mechanism on a cell membrane: (a) osmotic unbalance, (b) swelling, and (c)
membrane rupture.
fully clarified. Two theories of microbial inactivation have been proposed, out of which one is dielec-
tric breakdown theory (Zimmermann et al., 1976) and another is electroporation theory (Castro et al.,
1993). When suspension of cell is exposed to an electric field, it causes a trans-membrane poten-
tial and changes the membrane conductivity and permeability. When this induced trans-membrane
potential goes beyond the critical value (Ec), in many cellular systems membrane rupture occurs,
resulting in microbial inactivation (Figure 6.1) (Zimmermann et al., 1974; Coster and Zimmermann,
1975; Zimmermann, 1986; Castro et al., 1993). Another theory, i.e., theory of electroporation, can
also explain theory of dielectric breakdown (McLellan et al., 1991), which is based on the formation
of pores in the lipid or protein of the cell membrane. Due to the electric charges of lipid molecules
and permeability of lipid bilayer to ions, the cell membrane is susceptible to the applied electric
field. Lipid molecules reorient themselves due to electric charges under high electric field, thus
creating pores and damaging the bilayer barrier against ions (Figure 6.2). Except these, osmotic
effect and physiological impact may also influence the efficiency of electroplasmolysis (Weaver and
Chizmadzhev, 1996).
6.4 pEF EQUIpMENt
The equipment consists of a high-voltage pulse generator and a treatment chamber with a suitable
fluid handling system and necessary monitoring and controlling devices (Figure 6.3). Food prod-
uct to be treated is placed in the treatment chamber between two electrodes which are connected
together with a nonconductive material to avoid electrical flow from one to the other. High-voltage
electrical pulses are applied to the electrodes, which then conduct the high-intensity electrical pulse
to the product. Depending on the conductivity of food materials, which varies as a function of ions
present in the material, current conduction will take place. The food product experiences a force
per unit area, i.e., electric field, which is responsible for the reversible or irreversible damage of cell
membrane of the microorganisms.
Figure 6.3 block diagram of PeF set up.
6.4.1 treatment Chamber
The treatment chamber is one of the most important components of the PEF system. In the treat-
ment chamber food is exposed to the electric field pulses, consisting of at least two electrodes, one
on high voltage and the other on ground potential. The electrodes are separated by insulating mate-
rial in different geometric configurations. The materials selected to build up the treatment chamber
have to be washable (cleaning in place) or autoclavable and chemically inert with respect to foods.
The electrode and insulation material have to be food-grade and autoclavable, and furthermore their
electrochemical properties have to be considered. Stainless steel has been extensively used for the
electrodes. Other inert material includes carbon, gold, platinum, and metal oxides. Insulating mate-
rials used are polythene, polypropylene, nylon, polysulphone, plexiglass, and polyvinyl chloride.
Treatment chambers can be static (batch) or continuous (Figure 6.4). Static chambers are mostly
used in laboratory-scale studies, whereas continuous chambers are more suitable for pilot plant and
industrial-scale applications. The electrodes in the treatment chamber are arranged in different
geometric configurations: parallel plates, coaxial, and co-linear. Parallel plate systems are most
commonly used in batch mode while others are used in continuous flow operations. Parallel plates
provide the most uniform electric field in a large working area between the plates, but the treat-
ment intensity is reduced in boundary regions. Small volumes of treatment media are required and
treatment temperature is easy to maintain by cooling the electrodes and by slow repetition rates.
Moreover, the pulse number for each volume element is well known.
6.4.2 types of pulses
In PEF, high-intensity pulses of the range of 10–80 kV/cm are applied as short duration pulses
of around some micro to milliseconds duration using proper pulse forming networks (PFN). The
pulses used can be exponentially decaying, bipolar, square/rectangular, or oscillatory in nature.
The schematic representation of types of pulses is shown in Figure 6.5.
6.4.3 pulse Characteristics
The effect of PEF on the food product will be a function of pulse energy and pulse duration. Out
of the different types of pulses, the oscillatory type has the least efficiency level as it hinders the
Figure 6.4 Different configuration of electrodes (a) Parallel, (b) coaxial, and (c) co-linear for the treatment
chamber of PeF system. (From toepfl, s. et al., overview of pulsed electric field processing for
food, in: Emerging Technologies for Food Processing, pp. 93–108, sun, D. ed., elsevier Academic
Press, London, uK, 2005.)
(a) (b)
(c) (d)
Figure 6.5 schematic representation of types of pulses: (a) exponentially decaying pulse, (b) square wave
pulse, (c) bipolar pulse, and (d) oscillating pulse.
bacterial cells in liquid food from being exposed to high-intensity electrical field continuously. The
exponential type of pulse has a very high voltage surge but a very slow decay rate causing the long
tail section to produce heating but not enough to kill the bacteria in the food product. The square/
rectangular wave is much better than the other two categories as it can produce stable peak voltage
for elongated time interval, which will have an effective impact on the bacteria in the product.
6.4.4 pulse Generation Networks
Two types of pulse generation techniques widely reported are the conventional and power
electronics-based techniques.
6.4.4.1 Conventional Techniques
In conventional techniques, a high-voltage transmission line could produce a square/rectangular

wave pulse if the load impedance (offered by the food product) matches with the impedance of the
transmission line. But this may have a hindrance as the impedance of food product cannot be con-
trolled since it is an inherent capacity of the product. For this reason, PFN using transmission line
is not practically used and the other types of PFN used for different types of pulses are discussed
below.
Figure 6.6 shows a circuit to generate an exponential decay pulse. In this, a DC power source
is attached to a capacitor bank (C1 + C2 + C3) through a resistor Rc. A capacitor basically blocks
DC and, hence, gets charged towards the supply voltage. When the further path is closed through
Ignitron (a solid-state device with a very high current holding capability), the charge stored across
the capacitor bank finds a path to flow/discharge through a resistor combination of R1, R2 and the
resistance offered by the food product. R1 limits the current in case the sample sparks over and R2
controls the decay time if the food resistivity is higher than expected. The decay time of an RC cir-
cuit is given by τ = RC, where R is the total resistance offered in the decay path and C is the total
capacitance in the circuit. The capacitor charges/discharges completely in 5τ timing.
Figure 6.7 shows a circuit for generating bipolar pulses from a monopolar DC supply. The
capacitor C1 is charged using a DC power supply through Rc. When switch SW1 is closed, the voltage
across C1 is discharged through C2 and R L. During this discharge sequence a positive exponential
pulse is generated and C2 is also charged simultaneously with the polarity as indicated in Figure 6.7.
After T seconds SW1 and SW2 are closed and the latter acts as a bypass for R LC2 combination.
Figure 6.6 Circuit for generation of exponentially decaying pulses.

Figure 6.7 Circuit for generation of bipolar pulses from monopolar DC supply.
Figure 6.8 Circuit for generation of oscillatory pulses.
At this juncture the voltage across C2 will act as supply to the treatment chamber and this voltage
discharges across the food product. This pulse will be negative as −ve terminal of capacitor C2 is
connected to the treatment chamber.
Figure 6.8 shows the circuit for generation of oscillatory decaying pulses. In this circuit the treat-
ment chamber forms a part of the parallel LCR combination formed by the capacitor C, inductor L,
and resistor R2 in parallel with R1 and resistance of food material in treatment chamber. The capacitor
is charged as in other circuits through the charging resistor Rc. When the circuit is closed through the
ignitron switch, the capacitor begins to discharge and due to an LCR circuit being present, oscillations
occur depending on the value of inductor, capacitor, and resistor combination. The circuit must be oper-
ated in lightly damped condition for the oscillations to persist over a period of time (Qin et al., 1994).
6.4.4.2 Power Electronics Based Technique
With the advent of new power electronics-based high-speed devices a large number of power
electronics-based (electronic components working at higher voltages) PFNs are being proposed and
designed. One such circuit is shown in Figure 6.9. In this circuit the inductors of previous circuit
(shown in Figure 6.8) are replaced by transformers. The transformers provide current isolation,
which helps in circuit protection. The circuit shows a bidirectional flyback converter capable of
producing narrow bidirectional exponentially decaying pulses. Bipolar pulses help in reduction
of particle deposition. This circuit comprises of metal oxide semiconductor filed effect transistor
Figure 6.9 bidirectional Flyback converter circuit.
(MOSFET, represented by M in circuit), DIACs (D), and transformers (T), and the food material
acts as the load resistance R L (Jambari et al., 2011).
There are some of the more complicated circuits for the production of PFN by certain modifi-
cations of the basic circuit. There are many literatures referring to the different types of lab-scale
PFNs based on MOSFET power supplies and others.
6.5 FaCtOrS CONtrIBUtING tO thE EFFICaCY OF pEF tEChNOLOGY
The factors contributing to the effectiveness of PEF technology can be grouped as technologi-
cal, biological, and media factors. Some of these critical factors include the electric field strength,
treatment time or duration of treatment, treatment temperature, pulse shape, type of microorganism,
growth stage of the microorganism, and characteristics of the treatment substrate.
6.5.1 technological Factors
6.5.1.1 Electric Field Strength (E)
Microbial inactivation increases with an increase in the E above a critical electric field strength
(Ec). Electric field strength is calculated as
V (6.2)
E=
D
where V is the voltage across the food material, D is the distance between the electrodes, and Ec
is highly dependent on cell size as well as cell orientation in the field. With decrease in cell size
the required field strength also increases, and variations in cell shape also result in a consider-
able increase of E. As it is clearly indicated by Equation 6.2 that E is inversely proportional to
D, i.e., with increase in the distance between the electrodes, a higher voltage is required to obtain
the desired E (i.e., Ec corresponding to the membrane breakdown).
6.5.1.2 Duration of Treatment
Total time or duration of treatment is calculated using Equation 6.3
t = nτp (6.3)
where n is the number of pulses applied and τp is the pulse width.

The treatment time increases either with the number of pulses or with the pulse duration.
However, it is important to observe that increasing the number of pulses, the total energy consump-
tion also increases. Moreover, long treatment times lead to temperature rise in the food samples.
6.5.2 Biological Factors
The susceptibility of the microorganisms to PEF inactivation is highly dependent on the mor-
phological and physiological characteristics of treated microorganisms such as cell type, cell size
and shape, cell density, the cell arrangement in suspension, and dielectric breakdown. Generally,
gram-positive vegetative cells are more resistant to PEF than gram-negative bacteria due to the
presence of rigid teichoic acid in its cell. Bacterial spores and viruses are not affected by the PEF
treatment (Lelieveld et al., 2007). PEF inactivation rate does not only vary for different species but
also for different growth phase characteristics of each species. In the life cycle of a microorganism,
cells in proliferating phase are less resistant to PEF than in stationery and lag phases (Saulis, 2010).
The cells with larger diameters, such as yeast, were killed at lower electric field than the cells
with smaller diameter (Napotnik et al., 2016). Yeast showed greater sensitivity to PEF treatments
than vegetative bacteria due to their larger size than most bacteria, and as a result they may exhibit
a lower breakdown trans-membrane potential. The efficiency of PEF inactivation is also dependent
on population of microorganisms in the food and the larger number of microbial cells indicated the
less effective PEF treatment.
6.5.3 Media Factors
Food is a complex matrix, constituting several components in different proportion. The com-
position of the substrate can have a significant effect on the response of microorganisms to PEF
treatments as cells stressed might revive in the presence of food matrix. It is not necessary that in
the broth and food, same type of reduction will be achieved while keeping all others condition like
microorganism, electric field intensity, etc., as it is believed that food may provide the protective
effect to a microorganism. Proteins, carbohydrates, lipids, and other food constituents can confer
a protective effect. So, keeping this in consideration we cannot extrapolate the studies of broth
directly into the food system at the industrial level. Microorganism in buffers and microbiological
media are more pressure sensitive as compared to foods. Hence, inactivation data obtained using
buffers or laboratory media should not be extrapolated to real food situations where more severe
treatments may be needed to achieve the same level of inactivation.
Fluid (food product matrix) properties, such as electrical conductivity, ionic strength, and pH
strongly influence the PEF sensitivity of the microorganisms (Ortega-Rivas, 2011; Gamli, 2014).
The medium conductivity is an important parameter in PEF treatment and its correlation with
microorganism inactivation has been extensively investigated by several researchers (Marselles-
Fontanet et al., 2009; Buckow et al., 2013; Mohammed et al., 2016). Microbial inactivation is
enhanced at the lower ionic strength and conductivity (Krassowska and Filev, 2007; Saulis, 2010).
Vega-Mercado et al. (1996) reported that ionic strength was responsible for electroporation and
compression of the cells. The PEF treatment is more effective in a medium with lower conductivity
due to a larger difference in the ionic concentration between the suspension and the cell cytoplasm
(Shamsi and Sherkat, 2009). This large ionic gradient facilitates an increase in ionic substances
across the cell membrane, which weakens the membrane structure and makes it more vulnerable
to the PEF (Barsotti and Cheftel, 1999; Shamsi and Sherkat, 2009). The foods with high electrical
conductivity are difficult to process by PEF as they generate low peak electric fields across their
treatment chambers due to the high current (Stoica et al., 2011).
The influence of pH on the microbial inactivation has been documented in literature; however,
its relationship with PEF is not fully explored. In some studies, pH was reported to have no effect
on the microbial PEF inactivation (Ravishankar et al., 2002; Alvarez et al., 2003). However, in con-
trast, some studies reported lower microbial resistance at neutral pH (Alvarez et al., 2000; Geveke
and Kozempel, 2003) or in acidic medium (Saldana et al., 2011) to PEF treatment.
6.6 KINEtIC MODELS FOr pEF prOCESSING
Several studies have demonstrated the efficacy of PEF for inactivating a wide spectrum of
gram-negative and gram-positive bacteria in suspensions, as well as in liquid food items. The rate and
pattern of PEF-induced microorganism inactivation is quite variable and influenced by the processing
conditions, medium composition, and microorganism type/strain. Therefore, there is a need for
accurate prediction of the inactivation behaviour of food-borne microorganisms, as well as accurate
characterization of their resistance to PEF. Kinetic modelling is helpful for determining the most effi-
cient processing parameters and for the prediction of the processing technique effects on microbial
inactivation and product shelf-life (Kaur and Rao, 2017). Although simple first-order-type inactivation
curves do sometimes occur for PEF-treated cells (Rodrigo et al., 2003; Amiali et al., 2007; Pina-Pérez
et al., 2009), significant deviations from linearity (such as sigmoidal curves, curves with shoulders, and
tailing) have been reported by several researchers (Zhong et al., 2005; Rivas et al., 2006; Perez et al.,
2007). Various kinetic models, such as first-order kinetic model, Hulsgeger’s, Fermi’s, Weibull, and the
Log–logistic models, proposed to describe the microbial inactivation by PEF are presented in Table 6.1.
6.7 appLICatION ON FOOD QUaLItY
6.7.1 Milk and Milk products
In recent years many researchers have worked on application of PEF on milk and milk products
to preserve the sensory and physico-chemical quality. Shamsi (2008) observed that PEF treatment
(38 kV/cm and 60°C at a flow rate of 60 mL/min) did not show any significant (p > 0.05) change to
casein micelle size. There were no substantial differences in the particle size after PEF treatment
compared to thermal pasteurization at 73°C and 30 s. PEF treatment in combination with mild
thermal treatment of whole milk with 35 kV/cm and pulse width of around 2.3 ms at 65°C extends
the shelf life of whole soy milk up to 24 days. Combined effect of these treatments improved energy
efficiency in the processing, reported by Sepulveda et al. (2009). PEF treatment (35 kV/cm and
2.3 μs at 65°C for less than 10 s) of HTST-pasteurized milk immediately after processing enhanced
the shelf life of milk up to 60 days and PEF treatment after eight days of HTST pasteurization of
milk enhanced the shelf life of milk up to 78 days (Sepulveda et al., 2005). Guerrero-Beltran et al.
(2010) conducted a research on application of PEF and thermal treatment to inactivate Listeria
innocua in whole milk and concluded that PEF treatment (30 and 40 kV/cm field strength, no of
pulses (1–30) at 20°C–71°C for less than 10 s) of whole milk resulted in a maximum of 4.3 log
table 6.1 Kinetic Models for pEF Microbial Inactivation
Maximum reduction
Kinetic Model pathogen Medium treatment Conditions (log CFU/mL) references
First-order Kinetics Enterobacter sakazakii rehydrated infant 10–40 kV/cm, pulse width 2.5 μs, treatment 1.2 Perez et al. (2007)
CeCt 858 formula milk time 360 μs, flow rate 1.8 L/h, 25°C
E. coli orange juice 15–40 kV/cm, pulse width 2.5 μs, 3.83 rivas et al. (2006)
treatment time 700 μs, flow rate
60 mL/min, <55°C
E. coli Phosphate buffer 50–280 V/cm, treatment time 0.5–25 min, Machado et al. (2010)
65 to 234 V, 25°C
Hulsherger and E. coli orange juice 15–40 kV/cm, pulse width 2.5 μs, 3.83 rivas et al. (2006)
niemann model treatment time 700 μs, flow rate
60 mL/min, <55°C
E. coli CGMCC 1.90 Carrot juice 5–25 kV/cm, pulse width 1.5 μs, 3.6 Zhong et al. (2005)
PuLseD eLeCtriC FieLD ProCessinG
treatment time 207–1449 s, flow rate

52.5 mL/min, <40°C
Log-logistic Salmonella Mcilvaine buffer 12–28 kV/cm, pulse width 1–15 s, pulse 6.5 raso et al. (2000)
senftenberg frequency 1–5 Hz, treatment time
0–600 μs, initial temperature 20°C
Bacillus cereus Mixed beverage the pulse width was fixed at 2.5 μs, the 4.0 Pina-Pérez et al. (2009)
of liquid whole flow rate was 30 mL/min, treatment
egg and skim milk times ranged from 60 to 3895 μs, and
the electric field strength was set at 15,
25, and 35 KV/cm., below 20°C
E. coli o157:H7 Apricot nectar 17–30 kV/cm, treatment times 0–210 μs, 4.19 evrendilek et al. (2013)
flow rate 50 mL/min, pulse duration
Listeria monocytogenes 3 μs, frequency 500 pps 3.82
Staphylococcus aureus 3.76
Weibull E. coli orange juice 15–40 kV/cm, pulse width 2.5 μs, 3.83 rivas et al. (2006)
treatment time 700 μs, flow rate
60 mL/min, <55°C
Salmonella senftenberg Liquid egg 20–45 kV/cm, square pulse width 3 μs, 3.3 Monfort et al. (2010)
775W treatment time 0–150 μs, 55°C
E. coli o157:H7 Apricot nectar 17–30 kV/cm, treatment times 0–210 μs, 4.19 evrendilek et al. (2013)
L. monocytogenes flow rate 50 mL/min, pulse duration 3.82
3 μs, frequency 500 pps
S. aureus 3.76
(Continued)
99
100
table 6.1 (Continued) Kinetic Models for pEF Microbial Inactivation

Maximum reduction
Kinetic Model pathogen Medium treatment Conditions (log CFU/mL) references
Fermi’s model Salmonella. Dublin skim milk 15–40 kV/cm, treatment time 12–127 μs, 4.0 sensoy et al. (1997)
(AtCC 15480) 10°C–50°C
E. coli CGMCC 1.90 Carrot juice 5–25 kV/cm, pulse width 1.5 μs, 3.6 Zhong et al. (2005)
treatment time 207–1449 μs, coaxial
treatment chamber, flow rate
52.5 mL/min, <40°C
Quadratic E. coli o157:H7 Apple juice 20–30 kV/cm, pulse width 5–125 μs, flow 3.6 saldana et al. (2011)
response model rate 3 L/h, <55°C
Salmonella enteritidis strawberry juice 35 kV/cm, bipolar pulse width 4 μs, flow 4.3 Mosqueda-Melgar et al.
rate 80–110 mL/min, <40°C (2008)
S. cerevisiae Grape juice 35 kV/cm, bipolar pulse width 5 μs, pulse 3.9 Marselles-Fontanet
frequency 303 Hz, flow rate 3.33 mL/s, et al. (2009)
inlet temperature 15°C, maximum
temperature <30.4°C
natural microflora Mango nectar 38 kV/cm, 70–120 Hz, pulse width 4.1 Kumar et al. (2015)
15–24 μs
natural microflora Passion fruit juice 38 kV/cm, 85–100 Hz, pulse widths 4.0 Kathiravan et al. (2013)
19–24 μs
non-tHerMAL ProCessinG oF FooDs
cycle reduction in microbial population. Li et al. (2008) conducted a study of inactivation of soy-
bean enzyme lipoxygenase with different pulse strength, time, pulse frequency, and pulse width
and revealed that maximum inactivation of lipoxygenase enzyme (88%) occurred at 42 kV/cm for
1036 μs with 400 Hz of pulse frequency and 2 μs of pulse width at 25°C.
A comparative study of PEF and thermal treatment on the thermodynamic components of milk
(milk fat, xanthine oxidase, caseins, and whey proteins) has been reported by Sharma et al. (2016).
PEF treatments did not affect temperatures of fat melting (T melting) or xanthine oxidase denatur-
ation (T denaturation) with electric field strength from 20 or 26 kV cm−1 for 34 μs with or without
pre-heating of milk (55°C for 24 s). On the other hand, thermal treatments of milk increased both ther-
modynamics components of milk (T melting of milk fat and the T denaturation for xanthine oxidase
by 2°C–3°C) and observed 13% less reduction of xanthine oxidase after PEF treatment in comparison
to thermal treatments. McAuley et al. (2016) conducted a comparative study of PEF and thermal treat-
ments on quality of raw, pasteurized, and pulse electric-treated milk. They observed no significant
difference for rennet ability between the treatments, recorded slight drop in pH after change in counts
and concentration of short chain volatiles acids derived from microbial activity like 2, 3-butanedione,
acetic acid, and other short chain free fatty acids derived from milk (e.g., butanoic and hexanoic
acids), following the trend of microbial activity in milk samples. Craven et al. (2008) compared effect
of PEF and UHT treatment on shelf life of milk after adding spoilage microorganism “Pseudomonas
isolates”. Less inactivation of Pseudomonas was observed at 15°C or 40°C in comparison to 50°C or
55°C heat treatment. Maximum inactivation (5 log reduction) of Pseudomonas was achieved by PEF
processing at 55°C with 31 kV cm−1 (139.4 kJ L−1). They concluded that maximum inactivation of
Pseudomonas in milk could be achieved with the addition of PEF treatment rather than heat treatment.
6.7.2 Fruits and Vegetable products
PEF helps to improve the unit operations during food processing. PEF improves osmo-dehy-
dration process by increasing mass transfer and evacuating some portion of the fruit’s original
electrolytes, influencing the functional properties of the cell homeostasis system (Traffano-Schiffo
et al., 2016). Parniakov et al. (2016) reported that osmodehydration combined with PEF resulted in
observable increase in the rate of the freezing/thawing process as compared to untreated samples.
Luengo et al. (2013) reported that PEF makes the extraction process better for polyphenols and
flavonoids from orange peel by pressing. PEF-treated samples pressurized for 30 min increased the
extraction yield for total polyphenols by 20%, 129%, 153%, and 159%, respectively, at 1, 3, 5, and
7 kV/cm field strength for 60 μs (20 pulses of 3 μs). PEF extraction process enhances the extract’s
antioxidant capacity with reducing time for extraction without using organic solvents.
Fruit pulp treated using moderate electric field strength exhibited high yield and better juice
characteristics. In comparison to pectinase and microwave heating, isorhamnetin 3-O-rutinoside
from fruit peels into juice was released in higher amounts due to PEF (Moussa-Ayoub et al., 2016).
Extraction of carotenoid from tomato peel using PEF at 5 kV/cm increased by 39% in comparison
with the control sample in hexane: ethanol: acetone mixture in ratio of 50:25:25 (Luengo et al., 2014).
Pulsed electric treatment during frying of Kumara sweet potato tubers results in reduced oil
content in fried kumara chips besides reduction in energy requirement for cutting and frying. PEF
with strength of 1.2 kV/cm resulted in 18% lower oil content and attainment of brown colour at low
temperature as compared to untreated samples when fried at 190oC (Liu et al., 2017).
PEF improve the quality of grapefruit juice in combination with sonication (Aadil et al., 2015).
PEF have been reported to maintain high carotenoid content (10%–20%) in tomato juices throughout
the storage than thermally and untreated juices (Vallverdú-Queralt, et al., 2013). Mild-intensity PEF
and high-intensity PEF treatments in combination could be used for production of tomato juices hav-
ing high carotenoid content and to maintain carotenoid throughout the storage. Contents of phenolic
compounds observed in PEF-treated apples stored at 4°C and 22°C for 48 h were higher than those of
untreated (Soliva-Fortuny et al., 2017). However, some studies suggested that there is no significant
effect of PEF on the physico-chemical properties of fruit juices. Cserhalmi et al. (2006) investigated
colour, pH, TSS, non-enzymatic browning index, viscosity, electric conductivity, organic acid con-
tent, volatile flavour compound, and hydroxyl-methyl-furfurol in citrus juices and did not observe
any significant difference in the physico-chemical properties of treated and untreated samples.
PEF treatment retained ascorbic acid by more than 74% (Moussa-Ayoub et al., 2017), in case
of PEF treatment of fruit beverages sweetened with Stevia rebaudiana and also enhanced the total
anthocyanins and carotenoids content (Carbonell-Capella et al., 2017). In comparison to heat-
pasteurized products, higher vitamin C content has been reported in high-intensity PEF-treated
orange juice and gazpacho. In orange juice and gazpacho, the vitamin C retained by 87.5%–98.2%
and 84.3%–97.1%, respectively, after PEF treatments (Elez-Martínez and Martín-Belloso, 2007).
High electric field strengths, pulse widths, and frequencies of treated watermelon juice showed higher
retention of lycopene and antioxidant capacity. However, severe high-intensity PEF treatments were shown
to reduce vitamin C. Maximal relative lycopene content (113%), antioxidant capacity retention (100%), and
vitamin C (72%) were shown in high-intensity PEF-treated samples at 35 kV/cm for 50 μs by using 7 μs
bipolar pulses at 200 Hz (Oms-Oliu et al., 2009). PEF-treated samples have more stability of flavonoids
and phenolic acids as compared to thermally pasteurized samples (Agcam et al., 2014).
PEF technology results in microbial inactivation thereby enhancing the shelf-life of food.
Timmermans et al. (2014) conducted a study to check effectiveness of the PEF system to inactivate
S. panama, E. coli, L. monocytogenes, and S. cerevisiae in apple, orange, and watermelon juices
at 20 kV/cm. S. cerevisiae was observed to be the most susceptible micro-organism, followed by
S. panama and E. coli. Increasing the electric field strength and pulse rise time resulted in decrease
of residual enzyme activity of polyphenol oxidase and peroxidase and major inactivation of these
enzymes was achieved by 35 kV/cm field strength and 2 μs-pulse rise time (Bi et al., 2013).
6.7.3 Other Food products
Various studies related to the implementation of PEF technology in solid food material have been
carried out in recent years (Keith et al., 1997; Gudmundsson and Hafsteinsson, 2001; Eshtiaghi and
Knorr, 2002; Jun et al., 2003; Han et al., 2009; Janositz et al., 2011; Han et al., 2012; Carbonell-Capella
et al., 2017). Liu et al. (2017) investigated the effect of PEF at 0.3–1.2 kV/cm having energy levels
ranging from 0.5 and 22 kJ/kg on the microstructure and frying features of sweet potato (kumara).
There was significant softening of the ground tissues noticed after PEF processing. Pre-treatment of
tubers with electric field strength of 1.2 kV/cm followed by frying at 190°C had shown 18% reduc-
tion of oil content compared to untreated samples. Gudmundsson and Hafsteinsson (2001) investi-
gated the impact of PEF as well as their combination with high pressure on microstructure of various
meat products, i.e., salmon, chicken, and lumpfish roes. PEF lower than 2 kV/cm and 20–40 pulses
reported to have remarkable influence in the reduction of muscle cells size. Effect of PEF treatment
was higher for salmon than the chicken samples, whereas roes seem to remain unaffected up to elec-
tric field strength of 18.6 kV/cm and 7 pulses. Combination of PEF and high pressure (200–300 MPa)
had more destructive effect on microstructure than application of PEF alone. According to Guderjan
et al. (2007), maximum permeabilization of the membrane was attained for hulled rapeseed 55%
at 7.0 kV/cm and 120 pulses (84 kJ/kg) and 17% at 5 kV/cm and 60 pulses (42 kJ/kg) for non-hulled
rapeseed. Application of PEF earlier before oil separation could enhance the oil yield along functional
food ingredient content (polyphenols, phytosterols, tocopherols, and antioxidants). Han et al. (2009)
reported that corn starch subjected to PEF with 50 kV/cm had shown lower gelatinization temperatures
and enthalpy value. Loss of granular shape and reduction in crystallinity degree as well as pasting
parameters (peak, breakdown, and final viscosity) were also reported after the treatment. Han et al.
(2012) found that PEF treatment of tapioca starch at energy levels greater than 49.36 J/g may destroy
the crystallinity of starch granules. There was a significant reduction in the gelatinization temperature,
enthalpy, and peak as well as breakdown viscosity with increasing PEF strength. In a study, it was
reported that PEF treatment of potato slices improved liberation of intracellular molecules from
permeabilized tissue along with low molecular substances uptake in the sample. PEF treatment was
reported to be responsible for the reduction in fat content during deep fat frying, suggesting its poten-
tial to be used in the production of French fries with low fat content (Janositz et al., 2011). According
to Belghiti and Vorobiev (2004), maximal juice yield with minimum energy consumption (6–7 kJ/kg)
can be obtained by subjecting the sugar beet to PEF at 940 Vcm−1 and 250 pulses. Keith et al. (1997)
reported that field strength, pulse width, polarity reversal, and pulse number affected the bactericidal
effect of PEF on microorganisms in flour. It has been reported that 0.6-log reduction of aerobic plate
counts population of dark rye flour could be achieved after the PEF strengths greater than 20 kV/
cm. Lebovka et al. (2004) reported that complete removal of tissue textural strength was not possible
through PEF treatment alone. Therefore, combined application of the mild heat and PEF is necessary
for achieving the additional softening of carrots, potatoes, and apples. For apples, the mild heat and the
PEF treatments allow for the disintegration of the tissue texture to the state of a freeze-thawed tissue.
6.8 CONCLUSIONS
During the last decades PEF processing received considerable attention due to its potential to
maintain safety and results in minimal changes in the sensory and nutritional properties of the
treated food. PEF treatment can be used as an alternative technique for preservation of foods. It has
potential to reduce S. panama, E. coli, L. monocytogenes, and S. cerevisiae in apple, orange, and
watermelon juices and the efficiency of the bactericidal effect is affected by the nature of the juice
as well as the treatment conditions. Pre-treatment of tubers with electric field before frying causes
reduction of oil content compared to untreated samples. PEF seems to be a promising method for
inactivation of various microorganisms for better food quality with safety.
rEFErENCES
Aadil, R.M., Zeng, X.A., Sun, D.W., Wang, M.S., Liu, Z.W., and Zhang, Z.H. (2015). Combined effects of
sonication and pulsed electric field on selected quality parameters of grapefruit juice. LWT-Food Sci.
Technol. 62(1): 890–893.
Agcam, E., Akyıldız, A., and Evrendilek, G.A. (2014). Comparison of phenolic compounds of orange juice pro-
cessed by pulsed electric fields (PEF) and conventional thermal pasteurisation. Food Chem. 143: 354–361.
Aguilera, J.M., Stanley, D.W., and Baker, K.W. (2000). New dimensions in microstructure of food products.
Trends Food Sci Technol. 11: 3–9.
Alvarez, I., Raso, I., Sala, F.I., and Condon, S. (2003). Inactivation of Yersinia enterocolitica by pulsed electric
fields. Food Microbiol. 20: 691–700.
Alvarez, I., Raso, J., Palop, A., and Sala, F.J. (2000). Influence of different factors on the inactivation of
Salmonella senftenberg by pulsed electric fields. Int J Food Microbiol. 55: 143–146.
Amiali, M., Ngadi, M.O., Raghavan, G.S.V., and Smith, J.P. (2006). Inactivation of Escherichia coli O157:
H7 and Salmonella enteritidis in liquid egg white using pulsed electric fields. J. Food Sci. 71: 88–94.
Amiali, M., Ngadi, M.O., Smith, J.P., and Raghavan, G.S.V. (2007). Synergistic effect of temperature and
pulsed electric field on inactivation of Escherichia coli O157: H7 and Salmonella enteritidis in liquid
egg yolk. J. Food Eng. 79(2): 689–694.
Barsotti, L., and Cheftel, J.C. (1999). Food processing by pulsed electric field. II. Biological aspects. Food
Rev. Int. 15(2): 181–213.
Belghiti, K.E.L., and Vorobiev, E. (2004). Mass transfer of sugar from beets enhanced by pulse electric field.
Food Bioprod. Process. 82(3): 226–230.
Bi, X., Liu, F., Rao, L., Li, J., Liu, B., Liao, X., and Wu, J. (2013). Effects of electric field strength and pulse
rise time on physicochemical and sensory properties of apple juice by pulsed electric field. Innov. Food
Sci. Emerg. Technol. 17: 85–92.
Buckow, R., Ng, S., and Toepfl, S. (2013). Pulsed electric field processing of orange juice: A review on micro-
bial, enzymatic, nutritional, and sensory quality and stability. Compr Rev. Food Sci. F. 12: 455–467.
Carbonell-Capella, J.M., Buniowska, M., Cortes, C., Zulueta, A., Frigola, A., and Esteve, M.J. (2017). Influence
of pulsed electric field processing on the quality of fruit juice beverages sweetened with Stevia rebaudi-
ana. Food Bioprod. Process. 101: 214–222.
Castro, A.J., Barbosa-Canovas, G.V., and Swanson, B.G. (1993). Microbial inactivation of foods by pulsed
electric fields. Food Process Preserv. 17: 47–73.
Coster, H.G.L., and Zimmermann, U. (1975). The mechanisms of electrical breakdown in the membrane of
Valonia utricularis. J Membr Biol. 22: 73–90.
Craven, H.M., Swiergon, P., Ng, S., Midgely, J., Versteeg, C., Coventry, M.J., and Wan, J. (2008). Evaluation
of pulsed electric field and minimal heat treatments for inactivation of pseudomonads and enhancement
of milk shelf-life. Innov. Food Sci. Emerg. Technol. 9: 211–216.
Cserhalmi, Z., Sass-Kiss, A., Tóth-Markus, M., and Lechner, N. (2006). Study of pulsed electric field treated
citrus juices. Innov. Food Sci. Emerg. Technol. 7(1): 49–54.
Elez-Martínez, P., and Martín-Belloso, O. (2007). Effects of high intensity pulsed electric field processing
conditions on vitamin C and antioxidant capacity of orange juice and gazpacho, a cold vegetable soup.
Food Chem. 102: 201–209.
Eshtiaghi, M.N., and Knorr, D. (2002). High electric field pulse treatment: Potential for sugar beet processing.
J. Food Eng., 52(3): 265.
Evrendilek, G.A., Altuntas, J., Sangun, M.K., and Zhang, H.Q. (2013). Apricot nectar processing by pulsed
electric fields. Int. J. Food Prop. 16: 216–227.
Gamli, F. (2014). Advance research in agriculture and veterinary science. Adv. Res. Agri. Vet. Sci. 1(2): 54–61.
Geveke, D.J., and Kozempel, M.F. (2003). Pulsed electric field effects on bacteria and yeast cells. J. Food
Process Preserv. 27: 65–72.
Guderjan, M., Elez-Martinez, P., and Knorr, D. (2007). Application of pulsed electric fields at oil yield and
content of functional food ingredients at the production of rapeseed oil. Innov. Food Sci. Emerg.
Technol. 8(1): 55–62.
Gudmundsson, M., and Hafsteinsson, H. (2001). Effect of electric field pulses on microstructure of muscle
foods and roes. Trends Food Sci. Technol. 12(3–4): 122–128.
Guerrero-Beltrán, J.Á., David, R.S., María, M.G-N., Barry, S., Barbosa-Canovas, G.V. (2010). Milk thermiza-
tion by pulsed electric fields (PEF) and electrically induced heat. J. Food Eng. 100(1): 56–60.
Han, Z., Zeng, X.A., Fu, N., Yu, S.J., Chen, X.D., and Kennedy, J.F. (2012). Effects of pulsed electric field
treatments on some properties of tapioca starch. Carbohyd. Polym. 89(4): 1012–1017.
Han, Z., Zeng, X.A., Zhang, B.S., and Yu, S.J. (2009). Effects of pulsed electric fields (PEF) treatment on the
properties of corn starch. J. Food Eng. 93(3): 318–323.
Jacob, H.E., Foster, W., and Berg, H. (1981). Microbial implication of electric field effects. II. Inactivation of
yeasts cells and repair of their cell envelope. Z Allg Mikrobiol. 21: 225–233.
Jambari, H., Azli, N.A., and Piah, M.A.M. (2011). Comparison of pulsed electric field generation techniques
for microbial inactivation application. J. Theor. App. Infor.Technol. 33(1): 22–31.
Janositz, A., Noack, A.K., and Knorr, D. (2011). Pulsed electric fields and their impact on the diffusion char-
acteristics of potato slices. Food Sci. Technol. 44(9): 1939–1945.
Jun, S., Irudayaraj, J., Demirci, A., and Geiser, D. (2003). Pulsed UV-light treatment of corn meal for inactiva-
tion of Aspergillus niger spores. Int J Food Sci Tech. 38(8): 883–888.
Kathiravan, T., Nadanasabapathi, S., and Kumar, R. (2013). Optimization of pulsed electric field processing
conditions for passion fruit juice (Passiflora edulis) using response surface methodology. Int J Adv. Res.
1(8): 399–411.
Kaur, B.P., and Rao, P.S. (2017). Modeling the combined effect of pressure and mild heat on the inactiva-
tion kinetics of Escherichia coli, Listeria innocua, and Staphylococcus aureus in black tiger shrimp
(Penaeus monodon). Front Microbiol. 8: 1311.
Keith, W.D., Harris, L.J., and Giffiths, M.W. (1997). Reduction of bacterial levels in flour by pulsed electric
fields. J. Food Process Eng. 21(1998): 263–269.
Krassowska, W., and Filev, P.D. (2007). Modeling electroporation in a single cell. Biophys. J. 92: 404–417.
Kumar, R., Bawa, A.S., Kathiravan, T., and Nadanasabapathi, S. (2015). Optimization of pulsed electric field
parameters for mango nectar processing using response surface methodology. Int. Food Res. J. 22(4):
1353–1360.
Lebovka, N.I., Praporscic, I., and Vorobiev, E. (2004). Effect of moderate thermal and pulsed electric field treat-
ments on textural properties of carrots, potatoes and apples. Innov. Food Sci. Emerg. Technol. 5(1): 9–16.
Lelieveld, H.L.M., Notermans, S., and Haan, S.W.H. (2007). Food Preservation by Pulsed Electric Fields
from Research to Application. Woodhead Publishing, Cambridge, UK.
Li, Y.Q., Chen, Q., Liu, X.H., and Chen, Z.X. (2008). Inactivation of soybean lipoxygenase in soymilk by
pulsed electric fields. Food Chem. 109: 408–414.
Liu, T., Dodds, E., Leong, S.Y., Eyres, G.T., Burritt, D.J., and Oey, I. (2017). Effect of pulsed electric fields on the
structure and frying quality of “kumara” sweet potato tubers. Innov. Food Sci. Emerg. Technol. 39: 197–208.
Luengo, E., Álvarez, I., and Raso, J. (2013). Improving the pressing extraction of polyphenols of orange peel
by pulsed electric fields. Innov. Food Sci. Emerg. Technol. 17: 79–84.
Luengo, E., Álvarez, I., and Raso, J. (2014). Improving carotenoid extraction from tomato waste by pulsed
electric fields. Front Nutr. 1: 1–12.
Machado, L.F., Pereira, R.N., Martins, R.C., Teixeira, J.A., and Vicente, A.A. (2010). Moderate electric fields
can inactivate Escherichia coli at room temperature. J. Food Eng. 96: 520–527.
Marselles-Fontanet, A.R., Puig, A., Olmos, P., Minguez-Sanz, S., and Martin-Belloso, O. (2009). Optimising
the inactivation of grape juice spoilage organisms by pulsed electric fields. Int. J. Food Microbiol.
130(3): 159–165.
McAuley, C.M., Singh, T.K., Haro-Maza, J.F., Williams, R., and Buckow, R. (2016). Microbiological and
physicochemical stability of raw, pasteurised or pulsed electric field-treated milk. Innov. Food Sci.
Emerg. Technol. 38: 365–373.
McLellan, M.R., Kime, R.L., and Lind, L.R. (1991). Electroplasmolysis and other treatments to improve apple
juice yield. J. Food Agri. 57: 303–306.
Mohammed, M.E.A., Amer Eissa, A.H., and Aleid, S.M. (2016). Application of pulsed electric field for micro-
organisms inactivation in date palm fruits. J. Food Nutr. Res. 4(10): 646–652.
Monfort, S., Gayan, E., Raso, J., Condon, S., and Alvarez, I. (2010). Evaluation of pulsed electric fields tech-
nology for liquid whole egg pasteurization. Food Microbiol. 27(7): 845–852.
Mosqueda-Melgar, J., Raybaudi-Massilia, R.M., and Martin-Belloso, O. (2008). Nonthermal pasteurization of
fruit juices by combining high-intensity pulsed electric fields with natural antimicrobials. Innov. Food
Sci. Emerg. Technol. 9(3): 328–340.
Moussa-Ayoub, T.E., Jaeger, H., Youssef, K., Knorr, D., El-Samahy, S., Kroh, L.W., and Rohn, S. (2016).
Technological characteristics and selected bioactive compounds of Opuntia dillenii cactus fruit juice
following the impact of pulsed electric field pre-treatment. Food Chem. 210: 249–261.
Moussa-Ayoub, T.E., Jäger, H., Knorr, D., El-Samahy, S.K., Kroh, L.W., and Rohn, S. (2017). Impact of pulsed
electric fields, high hydrostatic pressure, and thermal pasteurization on selected characteristics of
Opuntia dillenii cactus juice. Food Sci. Technol. 79: 534–542.
Oms-Oliu, G., Odriozola-Serrano, I., Soliva-Fortuny, R., and Martín-Belloso, O. (2009). Effects of high-
intensity pulsed electric field processing conditions on lycopene, vitamin C and antioxidant capacity of
watermelon juice. Food Chem. 115(4): 1312–1319.
Ortega-Rivas, E. (2011). Critical issues pertaining to application of pulsed electric fields in microbial control
and quality of processed fruit juices. Food Bioprocess Technol. 4: 631–645.
Ortega-Rivas, E. (2012). High Voltage Pulsed Electric Fields. In Non-Thermal Food Engineering Operations,
pp. 275–295. Springer, New York.
Parniakov, O., Bals, O., Lebovka, N., and Vorobiev, E. (2016). Effects of pulsed electric fields assisted osmotic
dehydration on freezing-thawing and texture of apple tissue PEF-treatment treatment chamber osmotic
dehydration apple juice glycerol. J. Food Eng. 183: 32–38.
Perez, P.M.C., Aliaga, R.D., Bernat, F.C., Enguidanos, R.M., and Lopez, A.M. (2007). Inactivation of
Enterobacter sakazakii by pulsed electric field in buffered peptone water and infant formula milk. Int.
Dairy J. 17(12): 1441–1449.
Pina-Pérez, M.C., Silva-Angulo, A.B., Rodrigo, D., and Martínez-López, A. (2009). Synergistic effect of
pulsed electric fields and CocoanOX 12% on the inactivation kinetics of Bacillus cereus in a mixed
beverage of liquid whole egg and skim milk. Int. J. Food Microbiol. 130: 196–204.
Qin, B-L., Zhang, Q., Barbosa-Cánovas, G.V., Swanson, B.G., and Pedrow, P.D. (1994). Inactivation of micro-
organisms by pulsed electric fields of different voltage waveforms. IEEE T Dielect El In. 1(6): 1047–1057.
Raso, J., Alvarez, I., Condon, S., and Trepat, F.J.S. (2000). Predicting inactivation of Salmonella senftenberg
by pulsed electric fields. Innov. Food Sci. Emerg. Technol. 1(1): 21–29.
Ravishankar, S., Fleischman, G.J., and Balasubramaniam, V.M. (2002). The inactivation of Escherichia coli
0157: H7 during pulsed electric field (PEF) treatment in a static chamber. Food Microbiol. 19: 351–361.
Rivas, A., Sampedro, F., Rodrigo, D., Martinez, A., and Rodrigo, M. (2006). Nature of the inactivation of Escherichia
coli suspended in an orange juice and milk beverage. European Food Res. Technol. 223(4): 541–545.
Rodrigo, D., Ruiz, P., Barbosa-Canovas, G.V., Martinez, A., and Rodrigo, M. (2003). Kinetic model for the
inactivation of Lactobacillus plantarum by pulsed electric fields. Int. J. Food Microbiol. 81(3): 223–229.
Saldana, G., Puertolas, E., Monfort, S., Raso, J., and Alvarez, I. (2011). Defining treatment conditions for
pulsed electric field pasteurization of apple juice. Int. J. Food Microbiol. 151(1): 29–35.
Saulis, G. (2010). Electroporation of cell membranes: The fundamental effects of pulsed electric fields in food
processing. Food Eng. Rev. 2: 52–73.
Sensoy, I., Zhang, Q.H., and Sastry, S.K. (1997). Inactivation kinetics of Salmonella Dublin by pulsed electric
field. J. Food Process Eng. 20(5): 367–381.
Sepulveda, D.R., Gongora-Nieto, M.M., Guerrero, J.A., and Barbosa-Canovas, G.V. (2005). Production of
extended-shelf life milk by processing pasteurized milk with pulsed electric fields. J. Food Eng. 67: 81–86.
Sepulveda, D.R., Gongora-Nieto, M.M., Guerrero, J.A., and Barbosa-Canovas, G.V. (2009). Shelf life of whole
milk processed by pulsed electric fields in combination with PEF-generated heat. Food Sci. Technol. 42:
735–739.
Shamsi, K. (2008). Effects of pulsed electric field processing on microbial, enzymatic and physical attributes
of milk and the rennet-induced milk gels. Ph.D. thesis. Melbourne, Australia: RMIT University.
Shamsi, S., and Sherkat, F. (2009). Application of pulsed electric field in non-thermal processing of milk. As.
J. Food Ag-Ind. 2(3): 216–244.
Sharma, P., Oey, I., and Everett, D.W. (2016). Thermal properties of milk fat, xanthine oxidase, caseins and
whey proteins in pulsed electric field-treated bovine whole milk. Food Chem. 207: 34–42.
Soliva-Fortuny, R., Vendrell-Pacheco, M., Martín-Belloso, O., and Elez-Martínez, P. (2017). Effect of pulsed
electric fields on the antioxidant potential of apples stored at different temperatures. Postharvest Biol.
Technol. 132: 195–201.
Stoica, M., Bahrim, G., and Cârâc, G. (2011). Factors that influence the electric field effects on fungal cells. Science
Against Microbial Pathogens: Communicating Current Research and Technological Advances. 291–302.
Timmermans, R.A.H., Groot, M.N.N., Nederhoff, A.L., Van Boekel, M.A.J.S., Matser, A.M., and Mastwijk,
H.C. (2014). Pulsed electric field processing of different fruit juices: Impact of pH and temperature on
inactivation of spoilage and pathogenic micro-organisms. Int. J. Food Microbiol. 173: 105–111.
Tina Batista Napotnik, T.B., Matej Reberšek, M., Vernier, P.T., Mali, B., and Miklavčič, D. (2016). Effects of high
voltage nanosecond electric pulses on eukaryotic cells (in vitro): A systematic rev. Bioelectrochem. 110: 1–12.
Toepfl, S., Shemer, C., Saldana-Navarro, G., and Heinz, V. (2005). Overview of pulsed electric field processing
for food. In: Emerging Technologies for Food Processing, pp. 93–108. Sun, D. Ed., Elsevier Academic
Press, London, UK.
Traffano-Schiffo, M.V., Tylewicz, U., Castro-Giraldez, M., Fito, P.J., Ragni, L., and Rosa, M.D. (2016). Effect
of pulsed electric fields pre-treatment on mass transport during the osmotic dehydration of organic
kiwifruit. Innov. Food Sci. Emerg. Technol. 38: 243–251.
Vallverdú-Queralt, A., Odriozola-Serrano, I., Oms-Oliu, G., and Lamuela-Raventós, R.M. (2013). Impact of
high-intensity pulsed electric fields on carotenoids profile of tomato juice made of moderate-intensity
pulsed electric field-treated tomatoes. Food Chem. 141: 3131–3138.
Vega-Mercado, H., Pothakamury, U.R., Chang, F.J., Barbosa-Cánovas, G.V., and Swanson, B.G. (1996).
Inactivation of Escherichia coli by combining pH, ionic strength and pulsed electric field hurdles. Food
Res. Int. 29(2): 117–121.
Weaver, J.C., and Chizmadzhev, Y.A. (1996). Theory of electroporation: A review. Bioelectroch Bioener. 41:
135–160.
Zhong, K., Chen, F., Wang, Z.F., Wu, J.H., Liao, X.J., and Hu, X.S. (2005). Inactivation and kinetic model for
the Escherichia coli treated by a co-axial pulsed electric field. Eur Food Res Technol. 221(6): 752–758.
Zimmermann, U. (1986). Electrical breakdown, electropermeabilization and electrofusion. Rev. Physiol.
Biochem. Pharmacol. 105: 175–256.
Zimmermann, U., Pilwat, G., and Riemann, F. (1974). Dielectric breakdown of cell membranes. Biophys.
J. 14: 881–899.
Zimmermann, U., Pilwat, G., Beckers, F., and Riemann, F. (1976). Effects of external electric fields on cell
membrane. Biophys. J. 14: 881–889.
ChaptEr 7
pEF processing of Fruits, Vegetables,

and their products
R. Kumar, S. Vijayalakshmi, T. Kathiravan, and S. Nadanasabapathi
CONtENtS
7.1 Introduction ......................................................................................................................... 107

7.2 Food Preservation Using Electric Field/Energy ................................................................. 108
7.3 Pulsed Electric Field Processing......................................................................................... 108
7.4 Principles of Pulsed Electric Field Processing ................................................................... 108
7.5 Pulsed Electric Field System .............................................................................................. 109
7.6 Pulsed Electric Field Processing Parameters...................................................................... 110
7.7 Microbial Inactivation of Vegetables and Fruits ................................................................. 111
7.8 Enzymes of Vegetables and Fruits ...................................................................................... 113
7.9 Textural Characteristics of Vegetables and Fruits .............................................................. 115
7.10 Rheological Properties of Vegetables and Fruits ................................................................ 117
7.11 Shelf Stability of PEF Processed Foods ............................................................................. 118
7.12 PEF as a Pre-treatment AID ............................................................................................... 119
7.13 Effect on Toxin Reduction in Vegetables and Fruits .......................................................... 119
7.13.1 Effect on Pesticide Residues................................................................................. 120
7.13.2 Effect on Toxic Microbial Metabolites................................................................. 120
7.14 PEF Processing at Defence Food Research Laboratory, Mysore ....................................... 121
7.15 Future Scenario ................................................................................................................... 121
References ...................................................................................................................................... 122
7.1 INtrODUCtION
Fruit and vegetables are an important component of a healthy diet and, if consumed daily in
sufficient amounts, could help in preventing major diseases. They are an imperative source of vita-
mins, minerals, antioxidative compounds, and dietary fibres. Consumption of vegetables, fruits,
and their products may prevent chronic non-contagious diseases, popularly referred to as civiliza-
tion diseases, such as ischemic heart disease, arterial hypertension, obesity, and diet-related cancer
(Kanai et al., 2006). Phytochemicals and antioxidants found in fruits and vegetables are capable of
neutralizing free radicals and play a major role in the prevention of certain diseases (Kaur and Kapoor,
2001). Health-promoting food at present encompasses products causing lowering of arterial blood
pressure, fruit and vegetable juices, low-fat dairy drinks, probiotics, supplemented with calcium,
dietary fibre, prebiotics, energy drinks, low-fat cheeses, and bread enriched with dietary minerals
107
(Urala and Lähteenmäki, 2007). In particular, vegetable and fruit juices are promising in creating
functional food products. Their advantages include natural colorants such as betalains (betacyanins
and betaxanthins) carotenoids, anthocyanins, and chlorophylls that are capable of neutralizing free
radicals responsible for the processes of degradation of cell structures and aberration at the level of
DNA (Kapadia et al., 1996; Stintzing and Carle, 2004). Herbs confine to the plant parts which are
valued for medicinal, aromatic, and savoury qualities. An herb is a natural component which cures
minor ailments or otherwise maintains better health. Generally, herbs are used at lower levels in
products for providing flavour as well as minor nutrients to impart better health benefits.
7.2 FOOD prESErVatION USING ELECtrIC FIELD/ENErGY
There are several different ways of employing electric energy for food pasteurization. These
include ohmic heating (Anderson and Finkelstein, 1919; Getchell, 1935; Palaniappan and Sastry,
1991), microwave heating (Bengtsson and Ohlsson, 1974; Balanis, 1989), low electric field stimula-
tion (Lawrie, 1985; Li et al., 1994), high-voltage arc discharge (Dunn and Pearlman, 1987; Mizuno
and Hori, 1988; Palaniappan et al., 1990; Jayaram et al., 1991), and high-intensity pulsed electric
field (HIPEF) application (Mertens and Knorr, 1992; Qin et al., 1994; Sitzmann, 1995).
7.3 pULSED ELECtrIC FIELD prOCESSING
Non-thermal food processing technologies have a promising application as an alternative to

thermal technologies, by improving safety whilst maintaining product quality and economic feasi-
bility. Each of the non-thermal technologies has its own specific applications in terms of the type of
food that has to be treated. Amongst all the non-thermal methods, pulsed electric field (PEF) is one
of the most promising methods for the inactivation of microorganisms, proving to be an alternative
for pasteurization of liquid foods. PEF has the potential to pasteurize several foods by the use of
high-voltage short pulses maintained at temperatures from 30°C to 40°C, having an effect similar
to pasteurization, yet without the thermal component. The basic definition of PEF technology relies
on the application of high-intensity PEFs (l0–80 kV/cm) for cell membrane disruption, which is
due to the induced electric fields causing perforation in microbial membranes by electroporation.
Induction of membrane potentials exceeding a threshold value often results in cell damage and
death (Zimmermann, 1986).
PEF is an emerging electro-pulse technology that has been established to be a suitable method for
food preservation, hence receiving the worldwide attention of food professionals. The main advan-
tage of this emerging technology is the retention of fresh quality attributes, which is well proven
in the preservation of fruit juices, purees, and sauces, as well as dairy and poultry liquid products,
wherein the pathogenic and spoilage flora, as well as some enzymes, are inactivated. Furthermore,
PEF seems to be a good alternative for increasing the yield during juice extraction, performing liquid
waste treatment, and controlling biofouling in cooling water systems (Toepfl, 2006).
7.4 prINCIpLES OF pULSED ELECtrIC FIELD prOCESSING
PEF technology is based on the pulsing power dispensed to the product held between a set
of electrodes, confining the gap of the treatment chamber. The PEF processing line comprises a
high-voltage pulse generator and a treatment (continuous or static) chamber with a suitable fluid
handling system and also the necessary monitoring and controlling devices and an aseptic packag-
ing system to avoid post-processing contamination (Figure 7.1).
PeF ProCessinG oF Fruits, VeGetAbLes, AnD tHeir ProDuCts 109
Figure 7.1 PeF unit operations layout. (From Vega-Mercado, H. et al., Pulsed electric fields in food preservations-
Chapter 33, in: Handbook of Food Preservation, 2nd ed., pp. 783–813, LLC, taylor & Francis Group,
boca raton, FL, 2007.)
PEF treatment chamber is continuous or static, where the positive and negative electrodes are
connected with a non-conductive material to avoid electrical flow from one to the other electrode.
The high-intensity electrical pulse was passed to the electrodes where the food products were placed
between the electrodes. The food item placed in between the electrode experiences a force per unit
charge, i.e., the electric field, responsible for the irreversible membrane breakdown in microorgan-
isms (Zimmermann and Benz, 1980). Inactivation of microorganisms exposed to high-voltage PEF
is because of the electro-mechanical instability of the cell membrane, whereas the dose of the appli-
cation is attuned by means of electric field intensity, total number of pulses, and treatment time. The
minimal processing time even during the continuous operational mode has made the implementa-
tion of the PEF system feasible and easy in the existing operating lines.
7.5 pULSED ELECtrIC FIELD SYStEM
The PEF processing unit encompasses a high-voltage power source, a series of capacitors for
storage of energy, a resistor to limit the charging current, a switch to discharge energy from the
capacitor across the food, and a treatment chamber. An oscilloscope is used to observe the pulse
waveform. The power source, a high-voltage DC generator, converts the voltage from a utility line
(110 V) into high-voltage AC, then rectifies to a high-voltage DC. To generate an electric field in the
food material for preservation, the stored energy from the capacitor is discharged through the treat-
ment chamber. The maximum voltage across the capacitor is equal to the voltage across the genera-
tor. The bank of capacitors is charged by a direct current power source obtained from amplified and
rectified regular alternative current main source. An electrical switch is used to instantaneously
discharge energy stored in the capacitor storage bank across the food held in the treatment chamber.
Apart from those major components, some adjunct parts are also necessary. High-voltage and high-
current probes are used to measure the voltage and current delivered to the chamber.
The efficiency and the effectiveness of the PEF system depends on treatment chamber con-
figuration; the process parameters like electric field strength, pulse characteristics (number, width,
polarity, length, and shape), treatment temperature (start and end), field gap, flow rate, frequency,
specific energy, and residence time; microbial characteristics like type of microbe, its growth phase,
initial load, medium composition, oxygen concentration, etc.; and also on product nature such as its
composition, conductivity, ionic strength, acidity, and water activity (Kumar et al., 2006).
7.6 pULSED ELECtrIC FIELD prOCESSING paraMEtErS
The successful application of PEF processing has been reported to be based on its processing
parameters, such as electric field intensity, shape and polarity of the pulse wave, total treatment
time, and total pulsing energy input. In the study by Heinz et al. (2003) and Toepfl et al. (2005), it
was reported that the critical field intensity needed for maximal microbial inactivation depends on
size and shape of the microbial cell and also on the orientation of electric field.
Pulse wave shape and polarity were vital factors affecting the microbial inactivation by PEF
system. Amongst the two commonly used pulse shapes (square and exponential), the exponen-
tial decaying pulses were reported to be more effective in the microbial inactivation. The bipolar
pulses were also reported to be the most appropriate pulse polarity to achieve maximal reduction in
microbial count (Qin et al., 1994; Beveridge et al., 2005; Evrendilek and Zhang, 2005).
The total time of treatment and pulse energy input has been reported to be the major reasons for
effective microbial reduction (Bendicho et al., 2003; Min et al., 2003). In the continuous treatment, the
velocity of the fluid depends on the radial coordinate of the chamber; hence, the treatment time also
becomes non-homogenous (Figure 7.2). Thus, the average of the maximum treatment time (at walls of
the pipe) and shortest treatment time (centre of the pipe) has to be considered to calculate the minimum
treatment time needed for the applications such as microbial inactivation (Meneses et al., 2011).
Figure 7.2 treatment time and velocity profile for a laminar fluid flow in a continuous collinear treatment
chamber. (From Meneses, n. et al., basics for modelling of pulsed electric field processing of
foods, in: Innovative Food Processing Technologies: Advances in Multiphysics Simulation,
1st ed., pp. 171–192, Knoerzer, K., Juliano, P., roupas, P., and Versteeg, C., eds., Wiley blackwell
Publishing, 2011.)
Alkhafaji and Farid (2007) established that there was a dynamic correlation between electrical
conductivity, total energy input, and temperature increase in the treatment chamber and it needs to
be monitored and controlled to achieve the desired quality with a minimal effect on the product.
The efficient PEF treatment, in general, was proved to be based on the electric conductivity
of the product to be treated. Though the salt content aids in better conduction of electric pulses,
the higher range of salt present makes it unsuitable for PEF processing as the higher electrical
conductivity reduces the electric resistance and creates the need for more energy and electric field
strength for effective treatment (IFT, 2000). The possible pasteurisation applications of PEF include
treatment of various products such as fruit juices, tomato products, beers and wines, liquid dairy
products, and egg products (Floury et al., 2006).
7.7 MICrOBIaL INaCtIVatION OF VEGEtaBLES aND FrUItS
PEF technology ventures into possibilities for being applied for processing and utilization of raw
products and reduction of microbial contaminants from food microbiota. PEF treatment has been
found to be efficient and effective against the vegetative cells of microorganisms in food products
without significant loss of flavour, colour, or nutrients. The yeasts show a higher sensitivity to elec-
tric fields than vegetative forms of bacteria due to their larger size. PEF has only been proven to be
effective on vegetative cells (Stoica et al., 2011).
In general, spores of bacterial and fungal origin tend to be more resistant to inactiva-
tion when treated in the dry state (Brandon and Neuhauser, 1978). The circumstances of
sporulation/post-sporulation, the re-hydration, and the age of the spores greatly affects their
physiological state and thereby influences their resistivity to heat, inhibitors, and their germinabil-
ity (Dantigny and Nanguy, 2009). Alvarez et al. (2006) reported that ascospores and conidiophores
were sensitive to PEF treatment. However, the degree of resistance of moulds or yeast ascospores
is not yet established (Stoica et al., 2011). Though the effect of PEF on microbes has been estab-
lished, its effect on toxins produced by the microbes is yet to be investigated.
The effect of inactivation of microbes has been investigated and reported by many authors:
Escherichia coli (Zhao et al., 2011; Timmermans et al., 2014), Listeria innocua (Guerrero-Beltrán
et al., 2010), Listeria monocytogenes (Timmermans et al., 2014), Staphylococcus aureus (Zhao et al.,
2008), Saccharomyces cerevisiae (Timmermans et al., 2014), other microbes (Timmermans
et al., 2014), and Bacillus subtilis spores (Siemer et al., 2014a, 2014b).
Qin et al. (1994) reported that a PEF treatment at 12 kV/cm with 20 exponential decay pulses
inactivated 4 log cycles of S. cerevisiae in apple juice. Harrison et al. (1997) observed that a PEF
treatment at 40 kV/cm reduced the concentration of S. cerevisiae, inoculated initially, in apple juice
from 8 × 107 CFU/mL to 4 × 104 CFU/mL. They showed transmission electron microscopy (TEM)
micrographs of PEF-treated S. cerevisiae in apple juice. The PEF treatment disrupted yeast cells
and resulted in the almost total absence of ribosome bodies. Raso et al. (1998) reported that 6 log
cycles of Byssochlamys fulva conidiospores in cranberry juice were inactivated by PEF treatment
at 36.5 kV/cm. Jin and Zhang (1999) reported that a PEF treatment at 40 kV/cm for 150 μs reduced
about 5 log cycles in the total aerobic plate count and the yeast and mould count in cranberry juice.
The PEF process prohibited the growth of yeasts and moulds in the cranberry juice during 14 d of
storage at 4°C.
Qiu et al. (1998) reported that a pilot plant scale PEF treatment at 29.5 kV/cm for 60 μs inacti-
vated aerobic microorganisms in reconstituted orange juice by 4.2 log cycles. In a similar study by
Min and others (2002), the commercial scale PEF treatment at 40 KV/cm for 97 μs resulted in an
inactivation of 6 log cycles of total aerobic plate count and the yeast and mould count in orange juice.
Jia et al. (1999) reported that the amount of the total aerobic plate count and the yeast and mould
count in freshly squeezed orange juice were reduced by a PEF treatment at 30 kV/cm for 240 μs.
The PEF treatment was observed to be as effective as the thermal treatment at 90°C for 1 min in
reducing the total aerobic plate count and the yeast and mould count in the single-strength orange
juice. The total aerobic plate count and the yeast and mould count of the PEF-treated orange juice
were < 1 CFU/ml at 4°C for 6 weeks. McDonald et al. (2000) inoculated Leuconostoc mesenteroi-
des, E. coli, and Listeria innocua into orange juice and treated the inoculated orange juice with PEF
at 30 kV/cm. They achieved more than 5 log reductions of L. mesenteroides, E. coli, and L. innocua
inoculated in orange juice. They also obtained a maximum 2.5 log reduction of S. cerevisiae asco-
spores in orange juice by a PEF treatment at 50 KV/cm. Researchers observed 7-log reduction in
the total aerobic plate count and the yeast and mould count in orange juice after a PEF treatment at
35 KV/cm for 59 μs.
Garcia et al. (2005) evaluated and compared the PEF resistance of four gram-positive (Bacillus
subtilis, Lactobacillus plantarum, Listeriamonocytogenes, Staphylococcus aureus) and four gram-
negative (Escherichia coli, E. coli O157:H7, Yersinia enterocolitica, Salmonella serotype senften-
berg 775W) bacterial strains under the same treatment conditions. The most PEF-resistant bacteria
depended on the treatment medium pH. L. monocytogenes, which showed the highest PEF resistance
at pH of 7.0, was the most sensitive at pH 4.0. The highly PEF-resistant strains at pH 4.0 were the
gram-negatives such as E. coli O157:H7 and S. senftenberg. A successive holding of PEF-subjected
cells at pH 4.0 for 2 h increased the degree of inactivation up to 4 extra log10 cycles depending on
the bacterial strain investigated.
Elez-Martinez et al. (2006) subjected orange juice to HIPEF treatment with bipolar 4-μs pulses
of 35 kV/cm strength for 1000 μs at 200 Hz to enhance its microbial shelf life when stored at 4°C
and 22°C. HIPEF treatment ensured the microbiological stability of orange juice stored for 56 days
under refrigeration, but spoilage by naturally occurring microorganisms was detected within
30 days of storage at 22°C. Mosqueda-Melgar et al. (2008) studied the effect of high-intensity PEF
combined with citric acid or cinnamon bark oil against populations of Escherichia coli O157:H7,
Salmonella enteritidis, and Listeria monocytogenes in melon and watermelon juices. Populations of
the respective microbes were reduced by more than 5.0 log10 CFU/mL in HIPEF-processed melon
(35 KV/cm for 1709 μs at 193 Hz and 4 μs pulse duration) and watermelon (35 KV/cm for 1682 ms
at 193 Hz and 4 μs pulse duration) juices containing 2.0% and 1.5% of citric acid, respectively, or
0.2% of cinnamon bark oil. These treatments were also able to inactivate mesophilic, psychrophilic,
mould, and yeast populations effectively, leading to a shelf-life of more than 91 days in both juices
when stored at 5°C.
Evrendilek et al. (2000) also studied the effect of bench scale PEF treatment on E. coli
O157:H7 in apple juice and cider. The microbial reduction of 4.5 log was said to be achieved when
subjected to treatment of 34 kV/cm for 166 μs. Similarly, Cserhalmi et al. (2002) studied the effect
of PEF treatment of the inactivation of S. cerevisiae and B. cereus (cells and spores) inoculated in
sterilized apple juice and saline solution (0.15% NaCl), respectively. The yeast count reduction was
reported to be dependent on the electric field strength and pulse number. A maximal reduction of
yeast (S. cerevisiae) was obtained when the inoculated apple juice was subjected to a treatment with
10.4 pulses of 20 kV/cm strength. The PEF treatment on the B. cereus cells was observed to result in
a reduction of 1 log when treated with 10.4 pulses of 20 kV/cm strength. This showed that B. cereus
cells were less sensitive to PEF treatment.
Eisa et al. (2003) investigated the effect of PEF on aflatoxin production of Aspergillus flavus in
culture medium and corn grains. The multiple exposures of A. flavus-inoculated PDA medium and
corn grains to electric fields of varying frequency and treatment time were reported to affect the fun-
gal growth and also its efficacy to produce aflatoxin. It was also reported that the treatment resulted in
a significant amount of changes in protein and carbohydrate contents of the treated grains. Evrendilek
et al. (2004) studied the PEF treatment effectiveness of PEF treatment on microbial inactivation
in beer, on sensory properties and detection of electrode material migration. Beer samples were
treated by PEF for the inactivation of natural flora and inoculated cultures of Saccaromyces uvarum,
Lactobacillus plantarum, Bacillus subtilis, Rhodotorula rubra, and Pediococcus damnosus.

Inactivation of microbes as induced by the PEF treatment was observed to be 0.5, 4.1, 4.3, 4.7, 5.8, and
4.8 log10 colony forming units/ml in the above microorganisms, respectively. A significant increase
in the amount of minerals such as Cr, Zn, Fe, and Mn ions in the beer samples was found after PEF
treatment, leading to a statistically significant degradation in flavour and mouth feel.
The use of the PEF technology as an alternative system of microbiological control in wineries
was evaluated by Puértolas et al. (2009). The PEF resistance of different wine spoilage microorgan-
isms such as Dekkera bruxellensis, Dekkera anomala, Lactobacillus plantarum, and Lactobacillus
hilgardii in both must and wine was investigated by applying treatments ranging from 16 to 31 kV/cm
and from 10 to 350 kJ/kg at 24°C. The non-exponential kinetics of inactivation by PEF of these
microorganisms in both products had been described by mathematical equations based on the
Weibull distribution. The most favourable treatment of 186 kJ/kg at 29 kV/cm had been established
which permitted to reduce 99.9% of the spoilage flora of the must and wine and thereby limit the
risk of alteration of these products by microorganisms of genera Brettanomyces and Lactobacillus.
The effect of PEF processing parameters (electric field strength, pulse frequency, and treat-
ment time) on a mixture of microorganisms (Saccharomyces cerevisiae, Lactobacillus plantarum,
Lactobacillus hilgardii Gluconobacter oxydans, and Kloeckera apiculata) present in grape juice
and wine was evaluated (Marsellés-Fontanet et al., 2009). Yeast and bacteria were inactivated
by the PEF treatments resulting in reductions that ranged from 2.24 to 3.94 log units and also
proved to be affected by PEF parameters. Optimal inactivation of spoilage microorganism mixture
was predicted by the RSM models at 35.0 kV/cm−1 with 303 Hz pulse width for 1 μs. Inactivation
was greater for yeasts than in bacteria, as predicted by the RSM. The utmost efficacy of the PEF
treatment for inactivation of microorganisms in grape juice was observed around 1500 MJ/L for all
the microorganisms investigated. The PEF pasteurization of pomegranate juice for the microbial
inactivation was studied by Guo et al. (2014) for improving its shelf stability. The PEF treatment was
performed at electric field strength of 35 and 38 kV/cm for 281 μs at 55°C and flow rate of 100 L/h.
The microbial analysis revealed that the reduction of the aerobic microorganism remained unaltered
till the end of storage study.
7.8 ENZYMES OF VEGEtaBLES aND FrUItS
Inactivation of enzyme in food processing is very important for food preservation. An appli-
cation of electric field to food leads to association or dissociation of protein functional groups,
changes in helices alignment, and movements of charge chains (Tsong and Astunian, 1986). Castro
et al. (2001) observed that PEF treatment at 22.3 kV/cm with a pulse width of 0.78 μs tended to
associate and aggregate alkaline phosphatase enzyme, while electrical charge applied on the dipole
on the enzyme polarized and led to aggregate. The main mechanism of the enzyme inactivation was
regarded as aggregation of the enzyme by polarization.
Elez-Martinez et al. (2006) studied the effects of HIPEF processing (35 kV/cm for 1,000 μs; bipolar
4-μs pulses at 200 Hz) on the microbial shelf life and quality-related parameters of orange juice
during storage at 4°C and 22°C and compared to traditional heat pasteurization (90°C for 1 min) and
an unprocessed juice. HIPEF treatment ensured the microbiological stability of orange juice stored
for 56 days under refrigeration, but spoilage by naturally occurring microorganisms was detected
within 30 days of storage at 22°C. Pectin methylesterase (PME) of HIPEF-treated orange juice was
inactivated by 81.6%, whereas heat pasteurization achieved a 100% inactivation. Peroxidase (POD)
was destroyed more efficiently with HIPEF processing (100%) than with the thermal treatment
(96%). HIPEF-treated orange juice retained better colour than heat-pasteurized juice throughout
storage, but no differences were found between treatments in pH, acidity, and Brix. Vitamin C
retention was outstandingly higher in orange juice processed by HIPEF fitting recommended daily
intake standards throughout 56 days storage at 4°C, whereas heat-processed juice exhibited poor
vitamin C retention beyond 14 days storage (25.2%–42.8%). The antioxidant capacity of both treated
and untreated orange juice decreased slightly during storage.
The effects of HIPEF processing (35 kV/cm for 1700 ms applying 4 μs pulses at 100 Hz in bipolar
mode) on colour, viscosity, and also on the enzymatic activities such as PME and PG in strawberry
juice were studied by Aguilo-Aguayo (2009) and compared to those of heat treatments (90°C for
30 or 60 s) through 63 days of storage. L* and viscosity values of the HIPEF-processed juices were
higher than those found in the thermally processed juices. In addition, HIPEF-treated juice exhib-
ited lower 5-(hydroxymethyl)-2-furfural (HMF) concentration and browning index than heat-treated
juices during storage. In contrast, HIPEF-treated juice maintained low residual PME activity (13.1%)
for 63 days, whereas in the case of the thermally treated, 22.2% and 48.8% was retained after 60 s and
30 s, respectively. Strawberry juice treated by HIPEF achieved lower residual polygalacturonase
(PG) activity (73.3%) than those of heat-processed at 90°C for 60 s (76.2%) or 30 s (96.8%).
The aliphatic amino acids are the backbone of the globular structure of the alkaline phosphatase.
The PEF treatment preferably increases the accessibility of aliphatic hydrophobic region and it moved
from the hydrophilic surface of alkaline phosphatase, resulting in phosphatase unfolding. This also
suggested another possible mechanism of the inactivation of alkaline phosphatase (Castro et al.,
2001). The papain inactivation was due to loss of α-helical structure of enzyme and circular dichro-
ism (CD) analysis also demonstrated the differences in secondary structures of PEF-treated and
untreated papain. On the other hand, papain active site oxidation is not the major cause of inactiva-
tion (Yeom et al., 1999). Horseradish peroxidase (HRP) and pectinesterase (PE) enzymes are widely
present in the vegetable and fruit juices. PEF treatment (207 pulses of 25 kV/cm) was applied using
chambers with paralleled plane electrode and coaxial cylinder electrode. Results revealed that the
relative activity of HRP and PE activity was decreased 14.4% and 16.7%, 17% and 16.46% for the
parallel-plane chamber and the coaxial cylinder chamber, respectively.
PEF treatment effect on the POD and polyphenol oxidase (PPO) enzyme activity was observed
in buffered solution at 25 kV/cm for 1740 μs and PPO at 25 kV/cm for 744 μs. The relative residual
activity of POD and PPO decreased 32.2% and 76.2%, with the increase in electric field strength, treat-
ment time (Zhong et al., 2007). HIPEFs were utilized for inhibition of PME from tomato and a 93.8%
reduction of the initial activity was achieved with 400 pulses of 0.02 μs pulse-width at 24 kV/cm. The
classical exponential decay model as well as Hulsheger’s and Fermi’s empirical models adequately
described the enzymatic inhibition. Hulsheger’s equation yielded parameter values of 0.7 kV/cm,
0.48 μs, and 39 kV/cm, for critical electric field intensity, critical treatment time, and the independent
constant factor, respectively. For Fermi’s equation, the critical electric field and steepness parameters
ranged from 10.8 to 28 kV/cm and 5.7 to 11 kV/cm, respectively (Giner-Segui et al., 2000).
Freshly prepared apple juice was subjected to a combination of preheating to 50°C and a PEF
treatment time of 100 μs at 40 kV/cm for the inactivation of POD and PPO revealed that high-
est level of decrease was found. The PPO and POD enzymatic inactivation of 71% and 68% was
achieved, respectively. The enzyme inactivation was significantly higher than the conventional pas-
teurization, where the activity of PPO and POD activity decreased by 46% and 48%, respectively
(Riener et al., 2008). The higher inactivation of soybean lipoxygenase (LOX) (88%) was achieved,
when the soybean subjected to the PEF treatment of 42 kV/cm for 1036 μs with 400 Hz of pulse
frequency and 2 μs of pulse width at 25°C (Li et al., 2008). The residual activity of soybean LOX
decreased with the increase of treatment time, pulse strength, pulse frequency, and pulse width.
The inactivation of LOX and β-glucosidase (β-GLUC) in strawberry juice was subjected to
HIPEF. The residual activity of LOX remained at 65% and 70% in monopolar and bipolar mode,
respectively. In monopolar (61.6 Hz) and bipolar (218 Hz) low pulse frequencies, it did not have
adequate effect on LOX inactivation. On the other hand, higher β-GLUC inactivation was obtained
when the higher the pulse frequency and pulse width applied (Aguilo-Aguayo et al., 2008a).
PEF treatment with square wave and different combinations of pre-treatment temperature, electric
field strength, and treatment time were applied to the grapefruit juice and studied the inactivation
of PME and results revealed that PEF treatment (100 μs at 40 kV/cm) combined with preheating to
50°C achieved the highest level of PME inactivation (96.8%). Calculated D-values following a 50°C
preheat were 77.5, 76.0, and 70.4 μs at 20, 30, and 40 kV/cm, respectively. The activation energy
for the inactivation of PME by PEF was 36.2 kJ/mol (Riener et al., 2009). The effects of HIPEFs
on oxidative enzymes of fresh carrot juice was studied by Quitão-Teixeira et al. (2008) with mono
or bipolar mode, pulse width (from 1 to 7 μs), and pulse frequency (from 50 to 250 Hz). Response
surface methodology (RSM) was used to evaluate the effect of pulse POD inactivation on HIPEF
parameters. The electric field strength of 35 kV/cm and total treatment time of 1,000 μs at a tem-
perature below 35°C was used for the treatment and results revealed that 73% POD was inactivated,
when carrot juice subjected to 35 kV/cm for 1,000 μs, 6 μs pulse width at 200 Hz in bipolar mode.
HIPEF inactivation of POD, PME, and PG was studied by Aguilo-Aguayo et al. (2008b) and shows
that POD of was inactivated by 97%, whereas PME and PG inactivation was obtained 82% and 12%,
respectively.
Gazpacho (a Spanish ready-to-use cold vegetable soup) samples were exposed to 4 μs mono-
polar or bipolar square-wave pulses at 5–35 kV/cm electric field intensity for up to 1500 μs and
200 Hz of PEF treatment to inactivate the PE enzyme. Inactivation of PE was found to be greater
when treatment time and electric field intensity increased, and bipolar pulses were found to be more
effective inactivating PE than monopolar ones (Giner-Segui et al., 2009). The activity of sugar beet
o-diphenol oxidase and yeast invertase was studied by applying the PEF strength range between 3
and 60 kV/cm and pulse number variation from 1 to 30. The enzymes were found to be comparatively
resistant to PEF treatment irrespective of the electric pulses, conductivity, pH, and concentration
of their solutions. When the electric field strength ranged between 3 and 15 kV/cm applied to the
sample, the yeast invertase enzyme was reduced to only 10%–20% of initial activity. The activity
was decreased up to 30%, 90%, and 100%, when the electric field strength was increased to 17, 20,
and 60 kV/cm, respectively (Wawro et al., 2009).
7.9 tEXtUraL CharaCtErIStICS OF VEGEtaBLES aND FrUItS
PEFs have a targeted effect on biological tissue, where permeabilization of biological mem-
branes is achieved. The technique allows an accelerated mass transport in plant or meat tissue
and an improved release of intracellular substances (Toepfl and Heinz, 2010). In the tissues when
treated by pulse-electric field, the structure of the cells is altered due to two major changes: elec-
tropermeabilization and electroporation. The high-intensity electric pulses employed in the PEF
process induce reversible permeabilization in cells by pore developments in cell membranes (Knorr
et al., 2011). On further application of the electric field, the membranes can become permeable or
damaged (Lebovka et al., 2004). The electroporation of cell suspension and biological tissues was
influenced by orientation, size distribution, aggregation, arrangement of cells, localized cell den-
sity, solute concentration, and electric field distribution (Canatella et al., 2004; Toepfl et al., 2006;
Lebovka and Vorobiev, 2007; Pucihar et al., 2007; Pavlin et al., 2007).
Electrical breakage across the membrane created a critical transmembrane potential, altered
viscoelastic properties, and induced structural damages and compression of the membrane. This
dielectric breakdown of the cell membrane increased the permeability of the cell to electric cur-
rent and solutes (Bouzara and Vorobiev, 2000). The tissue damage caused by the PEF treatment
was reported to be minimal when compared to traditional heat treatments (Lebovka et al., 2004).
The irreversible damage caused by electric breakdown has found its application as pre-treatment to
drying and extraction process as well as to enhance mass transfer (Toepfl et al., 2006).
The PEF treatment-induced electro-permeabilization resulted in the softening of tissue and

loss of turgor. This mechanism makes it easy for the application as a pre-treatment to reduce
the cutting force of plant material. A continuous-mode PEF treatment for a short period and
low energy (~10 kJ/kg) resulted in the reduction of grinding energy or cutting force needed for
processing of plant or animal tissues. Loss of turgor pressure and elastic modulus was found to
change the cutting properties of the tissue. The extent of damage induced by PEF process depends
on the treatment protocol and also on the increase of the electric field strength and temperature
of treatment (Lebovka et al., 2005).
The application of PEF treatment on biological tissue can be used for induction of tissue dis-
integration, to improve mass transfer, and to induce desirable structural changes. The impact of
PEF on textural properties of potato tissue has been investigated (Figure 7.3). Fincan and Dejmek
(2003) and Lebovka et al. (2004) studied the effect of PEF on softening of apple, potato, and carrot
tissues and reported that there was reduction in elasticity modulus of the tissues. In another study
(Kraus, 2003), it was reported that cutting force reduced by 50% and cut quality of the sugar beet
was improved when subjected to PEF treatment.
During the PEF treatment, structural changes are influenced by the interaction between the
electromagnetic field and cellular material and thereby aid the juice extraction and tissue pressing
process. The various significant changes induced by the PEF treatment were reduction in perme-
ability (Galindo et al., 2008), increased diffusivity (Lebovka et al., 2007), enhanced drying (Wiktor
et al., 2013), inducing shrinkage on drying (Karathanos et al., 1996; Bazhal et al., 2002), alteration
in bulk and skeletal density (French, 1984), pore formation, and loss of turgor pressure.
Asavasanti et al. (2010) studied the influence of electric field strength and pulse number on
onion tissue integrity, texture, and chemistry. Their results suggested that there were two critical
electrical field strengths: one for the plasma membrane (67 V/cm, 10 pulses) and a higher one for the
tonoplast (200 V/cm, 10 pulses or 133 V/cm, 100 pulses).
Bazhal et al. (2003) proposed two investigational methods to determination effect of PEF treat-
ment on apple, potato, cucumber, aubergine, pear, banana, and carrot tissues. The two experimental
methods were estimation of time and total energy needed for inducing a characteristic damage and
estimation of the sample disintegration index caused by the input pulse energy. The study showed
Figure 7.3 Graphical representation of the impact of a PeF-treatment on the cutting force of potato tissue.
(From ranganathan, K. et al., Crit. Rev. Food Sci. Nutr., 2665–2694, 2016.)
that the electric field strength required to induce structural change was based on the tissue type and
also on the presence of secondary cell wall.
According to Galindo et al. (2008), the electropulsation of 100–500 V/cm slowed the diffu-
sion of dye through the cell wall of potato tissue. The decreased diffusion indicated the significant
decrease in permeability of cell wall at nanometer scale. The mechanism behind the reduction in
diffusion was the induced oxidative cross-linking, which reduced the pore dimensions and thereby
restricted the movement of particles through the apoplast. Structural collapse of the PEF-treated
samples was less severe when compared to the control untreated tissues (Bazhal et al., 2002). The
PEF pre-treated apple slices were observed to have significantly affected the bulk density, skeletal
density, porosity, pore size and volume shrinkage. The PEF pre-treated apple slices resulted in a
22% less volume-shrinkage than control samples (Wiktor et al., 2013).
The influence of combined PEFs and thermal treatment on the textural properties of apple
tissue and apple juice expression were studied by Lebovka et al. (2004). Force relaxation curves
were analysed for different regimes of PEF and thermal pre-treatment of apple tissue. The theory
of the effective relaxation time (α) was used to characterize the different modes of processing. Heat
treatment of apple tissue in its juice at a moderate temperature of 323 K for 10 min resulted in an
obvious softening of the tissues. The effect of mild thermal and PEF combination treatments was
shown to give more effective damage of apple tissue than the application of PEF treatment alone.
7.10 rhEOLOGICaL prOpErtIES OF VEGEtaBLES aND FrUItS
Rheology corresponds to the flow and deformation of a material under applied forces. It also
attempts to derive a relationship between the acting stress and the resulting deformation and/or
flow that takes place. These rheological properties have several applications in the field of food
processing, food handling, storage, and quality evaluation. Rheological data were required for
computation in many food processing operations which involve mixing, grinding, filtration and
extrusion etc. and also serve a significant role in the analysis of flow conditions in many food pro-
cessing operations such as pasteurization, concentration, dehydration including aseptic processing.
Rheological properties were determined generally by measuring shear stress and as a function of
shear rate and vice versa. The rheological analysis can be useful in new product development, qual-
ity control, determining the sensorial characteristics and also in designing and evaluation of new
process equipment (Ahmed et al., 2003; Augusto et al., 2012).
Aguilo-Aguayo et al. (2010b) studied the effects of HIPEF processing on colour, viscosity, and
related enzymes in watermelon juice during 56 days of storage and compared them to that of thermal
treatments. The high-intensity pulse electric pulse field-treated watermelon juice maintained brighter
red colour than that of thermal pasteurized juice and the application of HIPEF as well as thermal-
treated led to increase in viscosity of watermelon juice.
In general liquid foods were classified into Newtonian and non-Newtonian fluids based on their
rheological behaviour. The Newtonian behaviour corresponds to linear relationship between shear
stress and rate. Non-Newtonian fluids show a non-linear behaviour and can be represented with
various models namely Bingham, Herschel-Bulkley, Ostwald-De-Waele, Mizrahi-Berk and Casson
(Vandresen et al., 2009).
The rheological characteristics such as flow behaviour, viscosity, and other rheological parameters
such as consistency index etc. were affected by the PEF treatment. The authors Kumar et al. (2015)
studied the effect of PEF processing on the rheological behaviour of mango (Mangifera indica L.)
nectar. The rheological analysis was performed using a controlled stress rheometer. The PEF-treated
mango nectar behaved like a pseudo-plastic fluid and was found to obey the Herschel-Bulkley model
(0.9780 < r < 0.9999, p ≤ 0.01). The consistency coefficient and flow behaviour index of mango
nectar was said to be significantly affected by the PEF treatment.
7.11 ShELF StaBILItY OF pEF prOCESSED FOODS
The application of PEF processing is well known to increase the shelf stability of the prod-
uct with minimal effect on nutritional and sensorial attributes. It is in general said to retain the
natural “fresh-like” characteristics as the treatment involves a temperature at or below ambient
condition. Many researchers have studied the applicability of PEF system to extend the shelf sta-
bility of the products. The pilot plant scale PEF treatment at 35 kV/cm for 195 μs was reported
to decrease the numbers of the total aerobic plate count and yeast and mould count of cranberry
juice by more than 4 log cycles. The PEF-treated cranberry juice had the shelf life of 8 months
at 4°C, 37 days at 22°C, and 30 days at 37°C, respectively (Jin et al., 1998). In a similar study by
Sharma et al. (1998), it was reported that a pilot plant-scale PEF treatment at 32 KV/cm for 92 μs
reduced yeast and mould counts of whey protein-fortified orange juice from 1.4 × 105 CFU/mL to
<40 CFU/mL and made it microbiologically safe and stable for 5 months at 4°C. Similarly, Qiu
et al. (1998) and Min et al. (2002) studied the effect of PEF treatment on microbial inactivation in
orange juice and reported an extended shelf stability of 7 months at 4°C in aseptic packages and
196 days at 4°C, respectively.
The fresh apple cider and reconstituted apple juice were processed at 34 kV/cm for 94 μs in a
pilot-scale PEF system, packed and stored at three different temperature conditions (4°C, 22°C, and
37°C) for 70 days. During storage the treated samples were reported to retain their native quality
characteristics such as colour and vitamin C content. It was also stated that the treated products
possessed an improved microbial shelf life for a period of more than 67 days when stored in 4°C
and 22°C condition (Evrendilek et al., 2000).
Elez-Martinez et al. (2006) studied the effects of HIPEF processing (35 kV/cm for 1,000 μs; bipo-
lar 4-μs pulses at 200 Hz) on the microbial shelf life and quality-related parameters of orange juice
during storage at 4°C and 22°C and compared to traditional heat pasteurization (90°C for 1 min) and
unprocessed juice. HIPEF treatment ensured the microbiological stability of orange juice stored for
56 days under refrigeration, but spoilage by naturally occurring microorganisms was detected within
30 days of storage at 22°C. The quality attributes such as pH, acidity, Brix, vitamin C retention and
antioxidant capacity of the PEF-treated orange were reported to be minimally altered during the
storage period.
The application of HIPEF processing using bipolar pulses on watermelon juice was evaluated
by Aguilo-Aguayo et al. (2010a) as an alternative to conventional heat treatments (90°C for 30 or
90 seconds) in order to achieve better preservation of watermelon aroma compounds for 56 days of
storage at 4°C. HIPEF processing not only induced a rise (roughly 20%) in the hexanal, (E) -2- non-
enal, nonanal, 6-methyl-5-hepten-2-one and geranylacetone concentration, but also achieved less
reduction in the retention of volatiles than the thermal treatment at 90°C for 60 seconds. In contrast,
the content of (Z) -6- nominal, 1-nonanol, and (Z) -3-none-1-ol in the untreated and processed juices
remained unchanged. Even though a decrease in overall flavour compounds was observed dur-
ing storage irrespective of the treatment applied, HIPEF-treated juices retained better flavour than
heat-treated samples for at least 21 days of storage.
Marsellés-Fontanet et al. (2013) evaluated the influence of PEF and thermal technologies on
physical, chemical, microbiological, and nutritional properties of grape juice of different varieties.
The PEF treatment was observed to have a minimal effect on the quality attributes analysed when
compared to the thermal treatment. In another study, Guo et al. (2014) pasteurized pomegranate
juice (100% raw) using a commercial-scale PEF system. During the study, it was reported that the
maximal reduction in microbial count (total aerobic bacteria, yeast, and moulds) was obtained with
a minimal effect on the quality attributes such as TSS content, pH, sediment, total phenolic content,
antioxidant activity, anthocyanin content, and sensory properties. The stability of the treated juice
was reported to be achieved till 12 weeks of the storage study at 4°C when treated for 281 μs at 55°C
at electric field strength of 35–38 kV/cm and flow rate of 100 L/h.
7.12 pEF aS a prE-trEatMENt aID
A continuous mode PEF treatment for a short period and low energy (~10 kJ/kg) resulted in
the reduction of grinding energy or cutting force needed for processing of plant or animal tissues.
Loss of turgor pressure and elastic modulus was found to change the cutting properties of the tis-
sue. Dependent on process requirements, cell membrane permeabilization and loss of turgor can be
utilized for softening and modification of tissue, which may help in the replacement or improvisa-
tion of conventional processing techniques. The structural damage induced by the PEF process
depends on the treatment protocol and also increases with increase in electric field strength and
temperature of treatment (Lebovka et al., 2005).
According to Barba et al. (2015), the major advantages of PEF-assisted treatment were improved
mass transfer, enhanced extraction yield, reduction in processing time, decreased intensity of
extraction parameters, reduced degradation of heat-sensitive compounds, facilitated purifica-
tion extract, degradation of food contaminants, and reducing the energy costs and environmental
impact.
The influence of combined PEF and thermal treatment on the textural properties of apple tis-
sue and apple juice expression was studied by Lebovka et al. (2004). PEF-treatment requires rather
low power consumption, typically within 1–15 kJ/kg, and it defines the industrial attractiveness of
this technology. For sugar beet, many studies showed the efficiency of this technology and the tests
passed the industrial scale too (Vorobiev and Lebovka, 2010; Vidal, 2014). Industrial tests showed
that the implementation of PEF technology in the sugar beet industry enables important energy
savings which are greater than the implementation costs (Vidal, 2014). Recently, Mhemdi et al.
(2014) compared the impacts of thermal and (or) electrical pre-treatments of sugar beet cossettes on
the pressing yield, the energy consumption, and the qualitative characteristics of juice. The results
revealed that the raw juice expressed at 20°C with PEF showed higher purity, lower coloration, and
fewer colloids and proteins compared to the raw juices expressed at 50°C and at 80°C. In addition,
PEF-treated foods can be more easily purified by ultrafiltration than the filtrate of juice expressed
from the preheated sugar beet cossettes at 80°C.
The PEF-enhanced expression of the juice from slices of two different sizes was studied
for apple and carrot tissues (Praporscic et al., 2007). The experiments were carried out using a
laboratory filter-press cell and PEF-treatment system combo, operating at moderate electric field
strength (250–400 V/cm) and at constant pressure. It was shown that PEF-application of juice
expression from soft plant tissues allows for enhancement of the juice yields and to regulate quali-
tative characteristics of the juice. Enhancement of the juice expression (Y) after PEF-application
was accompanied with a noticeable decrease of absorbance (C) and increase of Brix value of
the juice for both apples and carrots. The PEF-application allows separating the second portion
of juice with the lowest absorbance and highest Brix values at yields 20%–30% for apple and 20%
for carrot.
Wiktor et al. (2013) studied the impact of PEF on the drying kinetics of apple tissue which were
treated at an intensity of 5–10 kV/cm and 10–50 pulse numbers and subsequently dried at 70°C
and air velocity of 2 m/s. PEF pre-treatment reduced the drying time up to 12% when treated with
50 pulses of field strength 10 kV/cm.
7.13 EFFECt ON tOXIN rEDUCtION IN VEGEtaBLES aND FrUItS
The samples exposed to the short electric pulses results in electroporation and electric break-
down, which results in the charging of the cell components and compounds/metabolites present in it.
The electric charging has been proved to have a significant influence on the components and metabo-
lites such as protein, vitamins, and toxic compounds such as pesticide residues, microbial toxins, etc.
7.13.1 Effect on pesticide residues
The PEF treatment of milk and other food products has caused conformational changes in the
proteins and other metabolites/compounds present in it. This led to the advancement of application
of PEF for the degradation of toxic pesticide residues present in food in trace amounts. The electric
charge/impedance produced during the processing were considered to contribute to the reduction of
pesticide residues. The pesticide reduction in various samples such as fruit juices, waste water, etc.
has been studied and reported by various authors.
A methodology for the reduction of organophosphorus pesticide in soybean milk by employing
HIPEF treatment has been patented by Zhijian et al. (2009). The soyabean milk containing meth-
amidophos, phorate, dimethoate, two cicada phosphorus, chlorpyrifos, and Mara parathion resi-
dues at lower concentration was degassed and pre-cooled before subjecting it to HIPEF with the
following parameters: pulse number of 26 and electric field strength of 8 kV/cm. The optimized
HIPEF treatment resulted in a degradation of 18.7%, 23.6%, 21.1%, 24.6%, 79.6%, and 52.8% of the
pesticides, respectively.
The electric field strength induced during PEF processing was reported to be responsible for
the degradation of pesticides like methamidophos, chlorpyrifos, diazinon, and dimethoate in apple
juice by Chen et al. (2009) and Zhang et al. (2012). Chen et al. (2009) hypothesised that higher volt-
age induces vibration and rotation of polar molecules which subsequently facilitates the degrada-
tion of pesticides. On the other hand, Zhang et al. (2012) reported that the hydrogen peroxide and
hydroxyl radicals formed might be the reason for pesticide degradation.
The fungicides-spiked white wine, on exposure to PEF of varying strength 5-20 kV/cm and
treatment time 0.5–2 ms, also resulted in the degradation of the fungicides (Delsart et al., 2016). The
concentrations of the studied fungicides (pyrimethanil, vinclozolin, cyprodinil, and procymidone)
were found to have significantly reduced after being subjected to PEF treatment. It was also reported
that sensitivity to PEF treatment varied with the fungicide and degradability was dependent on the
electric field strength and energy.
7.13.2 Effect on toxic Microbial Metabolites
The growth of microorganism during the post-harvest storage or during the processing results
in the production of toxic metabolites which are said to be highly stable and harmful to both
humans and animals. The PEF treatment has been established to be effective in the inactivation
of these microbes. The major problem faced worldwide is the contamination of fungal toxin in the
food products. The mycotoxins produced by the various fungal species are identified by IARC,
WHO, and FAO as highly thermostable and toxic, i.e., carcinogenic, toxigenic, etc. in nature. The
conventional method of removal of mycotoxin has been by physical screening for damaged or con-
taminated crops and various conventional thermal processing methods such as boiling, roasting,
cooking, etc. The thermal treatments were effective when the samples were treated at a higher
temperature or for a longer period, which causes the detrimental effect in the quality and sensory
attributes. The application of PEF treatment for the reduction of the toxin present has become a
boon to the food industry.
Eisa et al. (2003) has subjected the fungal organism (Aspergillus flavus)-inoculated potato dex-
trose agar (PDA) to PEF for various time periods and pulse frequencies. The exposure of fungal
culture to PEF showed a significant decrease in the aflatoxins production. Multiple exposure of
inoculated and non-inoculated corn grains showed that the aflatoxins (B1, B2, G1, G2, and total)
concentration was significantly lower than that of the control untreated sample.
The PEF process has a significant effect in the reduction and degradation of aflatoxins present
in food matrix. The thermal, PEF, and combination treatments were optimized using RSM for
maximization of reduction in spiked aflatoxin content in PDA using a batch chamber (Subramanian
table 7.1 products processed by pEF processing at DFrL, Mysore

Shelf Life (days)
S. No products at 5°C Under ambient Condition
1 tender coconut water 180 120
2 Flavoured milk 120 90
3 Mango nectar 180 120
4 Water melon juice 180 120
5 Musk melon juice 180 120
6 beetroot juice 180 120
7 Carrot juice 180 120
8 Passion fruit juice 180 120
9 Curd 180 21
10 tomato soup 180 120
11 Carrot soup 180 120
12 Mixed vegetable soup 180 120
13 Chicken clear soup 180 120
14 Mushroom soup 180 120
15 tender coconut water—nannari blend 180 120
et al., 2017; Vijayalakshmi et al., 2018). The HPLC analysis of the treated PDA revealed the PEF
process parameters pulse width (4–32 μs) and output voltage (20%–65%) had a significant effect
on the reduction on aflatoxins (B1, B2, G1, G2, and total) concentration. The optimized condition
for the reduction in aflatoxins was reported to vary with the pH of the PDA.
7.14 pEF prOCESSING at DEFENCE FOOD rESEarCh LaBOratOrY, MYSOrE
In India, DFRL, Mysore is the first research institute working on the PEF processing and its
combination with heat or sonication of pumpable liquid foods. DFRL has a continuous PEF sys-
tem. Pulse waveform, voltage, and intensity in the treatment chambers have been recorded with a
digital oscilloscope. The input process parameters such as output voltage, pulse width, frequency,
and flow rate and electrode gap can be set to suit the product. The institute has majorly contributed
to application of the PEF in processing of juices and soups for shelf life extension. Many products
were processed using the collinear continuous chamber individually or in combination and studied
for their quality and sensory attributes. The laboratory has worked extensively on the PEF process
optimization of liquid pumpable products and established the process parameters for the products
listed in Table 7.1; the final product of the same has been represented in Figure 7.4.
7.15 FUtUrE SCENarIO
The mechanism of PEF has still not been established completely. The advancement in analytical
techniques may help in solving the query on the mechanism by which the PEF acts. On the other side,
the application of PEF treatment is by default considered to be applicable for liquid (thin and thick)
matrix, though few studies have been conducted on solid matrices for the study on cell disintegration,
textural changes, pre-treatment, etc. The application of the process can be extended to solid matrix
by designing the geometric dimension of the treatment chamber as per the shape and structure of
the matrix. PEF treatment can also be established as an efficient physical tool in the reduction or
Figure 7.4 Products processed by PeF at DFrL, Mysore.
degradation of toxic compounds or metabolites, such as antibiotic residues and toxins inherited from
microbial, plants, and animals. The PEF treatment can be employed as pre-treatment in many food
processing industries, wherein the operations like extraction, drying, softening, etc. are involved. The
industrialization and indigenization of the PEF system are the immediate requirements, which are
still in the initial stages. The critical parameters involved in the application of PEF in indigenization
of products need an empirical approach, utilizing the advanced techniques and tools.
rEFErENCES
Aguilo-Aguayo, I., Sobrino-López, A., Soliva-Fortuny, R., and Martín-Belloso, O. (2008). Influence of high-
intensity pulsed electric field processing on lipoxygenase and β glucosidase activities in strawberry
juice. Innov Food Sci Emer Technol. 9: 455–462.
Aguilo-Aguayo, I., Soliva-Fortuny, R., and Martin-Belloso, O. (2008a). Comparative study on color, viscosity
and related enzymes of tomato juice treated by high intensity pulsed electric fields or heat. Eur Food
Res Technol. 227: 599–606.
Aguilo-Aguayo, I., Soliva-Fortuny, R., and Martin-Belloso, O. (2009). Avoiding non enzymatic browning by
high-intensity pulsed electric fields in strawberry, tomato and watermelon juices. J Food Eng. 92: 37–43.
Aguilo-Aguayo, I., Montero-Calderon, M., Soliva-Fortuny, R., and Martín-Belloso, O. (2010a). Changes
on flavor compounds throughout cold storage of watermelon juice processed by high-intensity pulsed
electric fields or heat. J Food Eng. 100: 43–49.
Aguilo-Aguayo, I., Soliva-Fortuny, R., and Martin-Belloso, O. (2010b). Color and viscosity of watermelon
juice treated by high-intensity pulsed electric fields or heat. Innov Food Sci Emerg Technol. 11: 299–305.
Ahmed, J., Ramaswamy, H.S., Alli, I., and Ngadi, M. (2003). Effect of high pressure on rheological character-
istics of liquid egg. LWT-Food Sci Technol. 36: 517–524.
Alkhafaji, S.R., and Farid, M. (2007). An investigation on pulsed electrics technology using new treatment
chamber design. Innov Food Sci Emerg Technol. 8: 205–212.
Alvarez, I., Condón, S., and Raso, J. (2006). Microbial inactivation by pulsed electric fields. In: Pulsed Electric
Fields Technology for the Food Industry, Fundamentals and Applications, Raso, J., and Heinz, V. Eds.,
Springer, New York.
Anderson, A.K., and Finkelstein, R. (1919). A study of the electropure process of treating milk. J Dairy Sci. 2: 374.
Asavasanti, S., Ersus, S., Ristenpart, W., Stroeve, P., and Barrett, D.M. (2010). Critical electric field strengths
of onion tissues treated by pulsed electric fields. J Food Sci. 75(7): E433–E443.
Augusto, P.E.D., Ibarz, A., and Cristianini, M. (2012). Effect of high pressure homogenization (HPH) on
the rheological properties of a fruit juice serum model. J Food Eng. 111: 474–477. doi:10.1016/j.
jfoodeng.2012.02.033.
Balanis, C.A. (1989). Advanced Engineering Electromagnetics. Wiley, New York.
Barba, F.J., Parniakov, O., Pereira, S.A., Wiktor, A., Grimi, N., Boussetta, N., Saraiva, J.A. et al. (2015).
Current applications and new opportunities for the use of pulsed electric fields in food science and
industry. Food Res Int. doi:10.1016/j.foodres.2015.09.015.
Bazhal, M., Lebovka, N., and Vorobiev, E. (2003). Optimisation of pulsed electric field strength for electroplas-
molysis of vegetable tissues. Biosys Eng. 86(3): 339–345.
Bazhal, M.I., Ngadi, M.O., and Raghavan, V.G.S. (2002). Textural changes in apple tissue during combined
pressure and pulsed electric field treatment. AIC 2002 Meeting. CSAE/SCGR Program. Saskatoon.
Saskatchewan. July 14–17. Paper No. 02–401.
Bendicho, S., Barbosa-Cánovas, G.V., and Martín, O. (2003). Reduction of protease activity in simulated milk
ultrafiltrate by continuous flow high intensity pulsed electric field treatments. J Food Sci. 68(3): 952–957.
Bengtsson, N.E., and Ohlsson, T. (1974). Microwave heating in food industry. Proc IEEE. 62(1): 44.
Beveridge, J.R., MacGregor, S.J., Marsili, L., Anderson, J.G., and Fouracre, R.A. (2005). The influence of
pulsed duration on the inactivation of bacteria using monopolar and bipolar profile pulsed electric fields.
IEEET Trans Plasma Sci. 33(4): 1287–1293.
Bouzrara, H., and Vorobiev, E. (2000). Beet juice extraction by pressing and pulsed electric fields. Int Sugar J.
102(1216): 194–200.
Brandon, J.R., and Neuhauser, K.S. (1978). Moisture effects on inactivation and growth of bacteria and fungi
in sludges. Sandia Laboratories, Albuquerque, NM, SAND78-1304, 1–24.
Canatella, P.J., Black, M.M., Bonnichsen, D.M., McKenna, C., and Prausnitz, M.R. (2004). Tissue electropora-
tion: Quantification and analysis of heterogeneous transport in multicellular environments. Biophysical J.
86: 3260–3268.
Castro, A.J., Swanson, B.G., Barbosa-Cánovas, G.V., and Zhang, Q.H. (2001). Pulsed electric field modification
of milk alkaline phosphatase activity. In: Electric Fields in Food Processing, pp. 65–82. Barbosa-Cánovas,
G.V., and Zhang, Q.H. Eds., Technomic, Lancaster, PA.
Chen, F., Zeng, L., Zhang, Y., Liao, X., Ge, Y., Hu, X., and Jiang, L. (2009). Degradation behavior of meth-
amidophos and chlorpyrifos in apple juice treated with pulsed electric fields. Food Chem. 112: 956–961.
Cserhalmi, Z., Vidacs, I., Beczner, J., and Czukor, B. (2002). Inactivation of Saccharomyces Cerevisiae and
Bacillus cereus by pulsed electric fields technology. Innov Food Sci Emerg Technol. 3: 41–45.
Dantigny, P., and Nanguy, S.P.-M. (2009). Significance of the physiological state of fungal spores. Int J Food
Microbiol. 134: 16–20.
Delsart, C., Franc, C., Grimi, N., de Revel G., Vorobiev, E., and Peuchot, M.M. (2016). Effects of
pulsed electric fields on four residual fungicides in white wines. In: 1st World Congress on
Electroporation and Pulsed Electric Fields in Biology, Medicine and Food & Environmental
Technologies (WC 2015), pp. 124–127. Jarm, T., and Kramar, P. Eds., IFMBE Proceedings, 53.
doi:10.1007/978-981-287-817-5_27.
Dunn, J.E., and Pearlman, J.S. (1987). Methods and apparatus for extending the shelf life of fluid food products.
U.S. patent 4,695,472.
Eisa, N.A., Ali, F.M., El-Habbaa, G.M., Abdel-Reheem, S.K., and Abou-El-Ella, M.F. (2003). Pulsed electric
field technology for checking aflatoxin production in cultures and corn grains. Egypt J Phytopathol
31(1–2):75–86.
Elez-Martinez, P., Soliva-Fortuny, R.C., and Martin-Belloso, O. (2006). Comparative study on shelf life of
orange juice processed by high intensity pulsed electric fields or heat treatment. Eur Food Res Technol.
222: 321–329.
Evrendilek, G.A., and Zhang, Q.H. (2005). Effect of pulse polarity and pulse delaying time on pulsed electric
fields-induces pasteurization of E. coli O157: H7. J Food Eng. 68: 271–276.
Evrendilek, G.A., Jin, Z.T., Ruhlman, K.T., Qiu, X., Zhang, Q.H., and Richter, E.R. (2000). Microbial safety
and shelf-life of apple juice and cider processed by bench and pilot scale PEF systems. Innov Food Sci
Evrendilek, G.A., Li, S., Dantzer, W.Q., and Zhang, Q.H. (2004). Pulsed electric field processing of beer:
Microbial, sensory and quality analyses. J Food Sci. 96(8): 228–232.
Fincan, M., and Dejmek, P. (2003). Effect of osmotic pretreatment and pulsed electric field on the viscoelastic
properties of potato tissue. J Food Eng. 59: 169–175.
Floury, J., Grosset, N., Leconte, N., Pasco, M., Madec, M., and Jeantet, R. (2006). Continuous raw skim
milk processing by pulsed electric field at non-lethal temperature: Effect on microbial inactivation and
functional properties. Lait. 86: 43–57.
French, D. (1984). Organization of starch granules. In: Starch: Chemistry and Technology. 2nd ed., pp. 183–247.
Whistler, R.L., Bemiller, J.N., and Paschall, E.F. Eds., Academic Press, New York.
Galindo, F.G., Vernier, P.T., Dejmek, P., Vicente, A., and Gundersen, M.A. (2008). Pulsed electric field reduces
the permeability of potato cell wall. Bioelectromagnetics. 29: 296–301.
Garcia, D., Gómez, N., Manas, P., Condon, S., Raso, J., and Pagan, R. (2005). Occurrence of sublethal injury
after pulsed electric fields depending on the microorganism, the treatment medium pH and the intensity
of the treatment investigated. J Appl Microbiol. 99(1): 94–104.
Getchell, B.E. (1935). Electric pasteurization of milk. Agric Eng. 16(10): 408.
Giner-Seguí, J., Elez-Martínez, P., and Martín-Belloso, O. (2009). Modelling within the Bayesian framework,
the inactivation of pectinesterase in gazpacho by pulsed electric fields. J Food Eng. 95: 446–452.
Giner-Seguí, J., Gimenoa, V., Espachsa, A., Elez-Martínez, P., Barbosa-Canovas, G.V., and Martin-Belloso,
O. (2000). Inhibition of tomato Licopersicon esculentum Mill. Pectin methylesterase by pulsed electric
fields. Innov Food Sci Emerg Technol. 1: 57–67.
Guerrero-Beltrán, J.A., Sepulveda, D.R., Góngora-Nieto, M.M., Swanson, B., and Barbosa-Cánovas, G.V. (2010).
Milk thermization by pulsed electric fields (PEF) and electrically induced heat. J Food Eng. 100: 56–60.
Guo, M., Jin, T.Z., Geveke, D.J., Fan, X., Sites, J.E., and Wang, L. (2014). Evaluation of microbial stability,
bioactive compounds, physicochemical properties, and consumer acceptance of pomegranate juice
processed in a commercial scale pulsed electric field system. Food Bioprocess Technol. 7: 2112–2120.
Harrison, S.L., Barbosa-Cánovas, G.V., and Swanson, B.G. (1997). Saccharomyces cerevisae structural
changes induced by pulsed electric field treatment. LWT-Food Sci Technol. 30: 236–240.
Heinz, V., Toepfl, S., and Knorr, D. (2003). Impact of temperature on lethality and energy efficiency of apple
juice pasteurization by pulsed electric fields treatment. Innov Food Sci Emerg Technol. 4(2): 167–175.
IFT. (2000). Special supplement: Kinetics of microbial inactivation for alternative food processing technologies—
IFT’s response to task order #1, US Food and Drug Administration: How to quantify the destruction kinetics
of alternative processing technologies. J Food Sci. 4: 108.
Jayaram, S., Castle, G.S.P., and Margaritis, A. (1991). Effects of high electric field pulses on L. brevis at
elevated temperatures. Proceedings of IEEE Industry Applications Society Annual Meeting. 647.
Jia, M., Zhang, Q.H., and Min, D.B. (1999). Pulsed electric field processing effects on flavour compounds and
microorganisms of orange juice. Food Chem. 65: 445–451.
Jin, Z.T., Ruhlman, K.T., Qiu, X., Jia, M., Zhang, S., and Zhang, Q.H. (1998). Shelf-life evaluation of pulsed
electric fields treated aseptically packaging material cranberry juice. IFT 1998 Annual Meeting Book of
Abstracts, Paper no.34B-10, 70.
Jin, Z.T., and Zhang, Q.H. (1999). Pulsed electric field inactivation of microorganisms and preservation of
quality of cranberry juice. J Food Process Eng. 23: 481–497.
Kanai, C., Pomerleau, J., Lock, K., and McKee, M. (2006). Getting children to eat more fruit and vegetables:
A systematic review. Prev Med. 42: 85–95.
Kapadia, G.J., Tokuda, H., Konoshima, T., and Nishino, H. (1996). Chemoprevention of lung and skin cancer
by Beta vulgaris (beet) root extract. Cancer Lett. 100(1–2): 211–214.
Karathanos, V.T., Kanellopoulos, N.K., and Belessiotis, V.G. (1996). Development of porous structure during
air drying of agricultural plant products. J Food Eng. 29(2): 167–183.
Kaur, C., and Kapoor, H.C. (2001). Antioxidants in fruits and vegetables: The millennium’s health. Int J Food
Knorr, D., Froehling, A., Jaeger, H., Reineke, K., Schlueter, O., and Schoessler, K. (2011). Emerging technolo-
gies in food processing. Ann Rev Food Sci Technol. 2: 203–235.
Kraus, W. (2003). The 2002 beet campaign – VDZ Zweigverein Süd. Zuckerindustrie. 128(5): 344–354.
Kumar, R., Bawa, A.S., Manjunatha, S.S., Kathiravan, T., and Nadanasabapathi, S. (2015). Effect of pulsed
electric field and pasteurisation treatments on the rheological properties of mango nectar (Mangifera
indica). Croat J Food Sci Technol. 7(1): 22–33. doi:10.17508/CJFST.2015.7.1.02.
Kumar, R., Sabapathy, S.N., and Bawa, A.S. (2006). Pulsed electric field processing of foods: A review.
Processed Food Industry. 9(10): 27–32.
Lawrie, R.A., (1985). Meat Science, 4th ed., Pergamon Press, New York.
Lebovka, N.I., Praporscic, I., Ghnimi, S., and Vorobiev, E. (2005). Temperature enhanced electroporation
under the pulsed electric field treatment of food tissue. J Food Eng. 69(2): 177–184.
Lebovka, N.I., Praporscic, I., and Vorobiev, E. (2004). Combined treatment of apples by pulsed electric fields
and by heating at moderate temperature. J Food Eng. 65: 211–217.
Lebovka, N.I., Shynkaryk, M.V., El-Belghiti, K., Benjelloun, H., and Vorobiev, E. (2007). Plasmolysis of sug-
arbeet: Pulsed electric fields and thermal treatment. J Food Eng. 80(2): 639–644.
Lebovka, N.I., and Vorobiev, E. (2007). The kinetics of inactivation of spheroidal microbial cells by pulsed
electric fields. E-print arXiv:0704.2750v1, 1–22.
Li, Y., Slavik, M.F., Griffis, C.L., Walker, J.T., Kim, J.W., and. Wolfe, R.E. (1994). Destruction of Salmonella
in poultry chiller water using electrical stimulation. Trans ASAE. 37(1): 211.
Li, Y.-Q., Chen, Q., Liu, X.-H., and Chen, Z.-X. (2008). Inactivation of soybean lipoxygenase in soymilk by
pulsed electric fields. Food Chem. 109: 408–414.
Marsellés-Fontanet, R.A., Puig-Pujol, A., Olmos, P., Mínguez-Sanz, S., and Martín-Belloso, O. (2009).
Optimising the inactivation of grape juice spoilage organisms by pulse electric fields. Int J Food
Microbiol. 130: 159–165.
Marsellés-Fontanet, R.A., Puig-Pujol, A., Olmos, P., Mínguez-Sanz, S., and Martín-Belloso, O. (2013). A com-
parison of the effects of pulsed electric field and thermal treatments on grape juice. Food Bioprocess
Technol. 6: 978–987.
McDonald, C.J., Lloyd, S.W., Vitale, M.A., Petersson, K., and Innings, F. (2000). Effect of pulsed electric
fields on microorganisms in orange juice using electric field strengths of 30 and 50 kV/cm. J Food Sci.
65(6): 984–989.
Meneses, N., Jaeger, H., and Knorr, D. (2011). Basics for modelling of pulsed electric field processing of foods.
In: Innovative Food Processing Technologies: Advances in Multiphysics Simulation, 1st ed., pp. 171–192.
Knoerzer, K., Juliano, P., Roupas, P., and Versteeg, C. Eds., Wiley Blackwell Publishing, IFT, USA.
Mertens, B., and Knorr, D. (1992). Developments of nonthermal processes for food preservation. Food
Technol. 46(5): 124.
Mhemdi, H., Bals, O., Grimi, N., and Vorobiev, E. (2014). Alternative pressing/ultrafiltration process for sugar
beet valorization: impact of pulsed electric field and cossettes preheating on the qualitative characteris-
tics of juices. Food Bioprocess Technol. 7: 795–805.
Min, S., Jin, Z.T., Yeom, H.W., Min, S.K., and Zhang, Q.H. (2002). Effects of commercial scale pulsed electric
field processing on the quality of orange juice. J Food Sci. 68(4): 1265–1271.
Min, S., Jin, Z.T., and Zhang, Q.H. (2003). Commercial scale pulsed electric field processing of tomato juice.
J Agri Food Chem. 51: 3338–3344.
Mizuno, A., and Hori, Y. (1988). Destruction of living cells by pulsed high-voltage application. IEEE Trans
Ind Appl. 24: 387.
Mosqueda-Melgar, J., Raybaudi-Massilia, R.M., and Martin-Belloso, O. (2008). Combination of high-intensity
pulsed electric fields with natural antimicrobials to inactivate pathogenic microorganisms and extend
the shelf-life of melon and watermelon juices. Food Microbiol. 25: 479–491.
Palaniappan, S., Richter, E.R., and Sastry, S.K. (1990). Effects of electricity on microorganisms: A review.
J Food Process Preserv. 14: 393–414.
Palaniappan, S., and Sastry, S.K. (1991). Electrical conductivity of selected juices: Influences of temperature,
solids content, applied voltage, and particle size. J Food Proc Eng. 14: 247.
Pavlin, M., Leben, V., and Miklavcic, D. (2007). Electroporation in dense cell suspension. Theoretical and
experimental analysis of ion diffusion and cell permeabilization. BBA. 1770(1): 12–23.
Praporscic, I., Shynkaryk, M.V., Lebovka, N.I., and Vorobiev, E. (2007). Analysis of juice colour and dry matter
content during pulsed electric field enhanced expression of soft plant tissues. J Food Eng. 79: 662–670.
Pucihar, G., Kotnik, T., Teissie, J., and Miklavcic, D. (2007). Electropermeabilization of dense cell suspensions.
Eur Biophys J. 36(3): 173–185.
Puértolas, E., López, N., Condón, S., Raso, J., and Álvarez, I. (2009). Pulsed electric fields inactivation of wine
spoilage yeast and bacteria. Int J Food Microbiol. 130: 49–55.
Qin, B.L., Zhang, Q., Barbosa-Cánovas, G.V., and Swanson, B.G. (1994). Inactivation of microorganisms by
pulsed electric fields of different voltage waveforms. IEEE Trans Dielectr Electr Insul. 1(6): 1047.
Qiu, X., Sharma, S., Tuhela, L., Jia, M., and Zhang, Q.H. (1998). An integrated PEF pilot plant for continuous
nonthermal pasteurization of fresh orange juice. Trans ASAE. 41(4): 1069–1074.
Quitão-Teixeira, L.J., Aguiló-Aguayo, I., Ramos, A.M., and Martín-Belloso, O. (2008). Inactivation of oxida-
tive enzymes by high-intensity pulsed electric field for retention of color in carrot Juice. Food Bioproc
Technol. 1: 364–373.
Ranganathan, K., Subramanian, V., and Shanmugam, N. (2016). Effect of thermal and nonthermal processing on tex-
tural quality of plant tissues. Crit Rev Food Sci Nutr. 56: 16, 2665–2694. doi: 10.1080/10408398.2014.908348.
Raso, J., Calderón, M.L., Góngora, M., Barbosa-Cánovas, G.V., and Swanson, B.G. (1998). Inactivation of
mold ascospores and conidiospores suspended in fruit. LWT-Food Sci Technol. 668–672.
Riener, J., Noci, F., Cronin, D.A., Morgan, D.J., and Lyng, J.G. (2008). Combined effect of temperature and pulsed
electric fields on apple juice peroxidase and polyphenol oxidase inactivation. Food Chem. 109: 402–407.
Riener, J., Noci, F., Cronin, D.A., Morgan, D.J., and Lyng, J.G. (2009). Combined effect of temperature and
pulsed electric fields on pectin methyl esterase inactivation in red grapefruit juice (Citrus paradisi).
Eur Food Res Technol. 228: 373–379.
Sharma, S.K., Zhang, Q.H., and Chism, G.W. (1998). Development of a protein fortified fruit beverage and its
quality when processed with pulsed electric field treatment. J Food Qual. 21: 459–473.
Siemer, C., Toepfl, S., and Heinz, V. (2014a). Inactivation of Bacillus subtilis spores by pulsed electric fields
(PEF) in combination with thermal energy – I. Influence of process- and product parameters. Food
Control. 39: 163–171.
Siemer, C., Toepfl, S., and Heinz, V. (2014b). Inactivation of Bacillus subtilis spores by pulsed electric fields
(PEF) in combination with thermal energy II. Modeling thermal inactivation of B. subtilis spores during
PEF processing in combination with thermal energy. Food Control. 39: 244–250.
Sitzmann, W. (1995). High voltage pulse techniques for food preservation. In: New Methods of Food
Preservation, p. 236. Gould, G.W. Ed., Blackie Academic & Professional, Chapman & Hall, New York.
Stintzing, F.C., and Carle, R. (2004). Functional properties of anthocyanins and betalains in plants, food and
in human nutrition. Trends Food Sci Technol. 15: 19–38.
Stoica, M., Bahrim, G., and Carac, G. (2011). Factors that influence the electric field effects on fungal cells.
In: Science Against Microbial Pathogens: Communicating Current Research and Technological
Advances, pp. 291–302. Mendez-Vilas, A. Ed., Formatex, Badajo, Spain.
Subramanian, V., Shanmugam, N., Ranganathan, K., Kumar, S., and Reddy, R. (2017). Effect of combi-
nation processing on aflatoxin reduction: Process optimization by response surface methodology.
J Food Process Preserv. 41(6): e13230. doi:10.1111/jfpp.13230.
Timmermans, R.A.H., Nierop Groot, M.N., Nederhoff, A.L., van Boekel, M.A.J.S., Matser, A.M., and
Mastwijk, H.C. (2014). Pulsed electric field processing of different juices: Impact of pH and tempera-
ture on inactivation of spoilage and pathogenic micro-organisms. Int J Food Microbiol. 173: 106–111.
Toepfl, S. (2006). Pulsed electric fields (PEF) for permeabilization of cell membranes in food- and bioprocessing
– applications, process and equipment design and cost analysis. Department of Food Process Engineering
and Food Biotechnology, Berlin University of Technology, Berlin, Germany. PhD thesis.
Toepfl, S., and Heinz, V. (2010). Role for pulsed electric fields. Industry innovation, The World of Food
Ingredients, p. 65.
Toepfl, S., Heinz, V., and Knorr, D. (2005). Overview of pulsed electric field processing for food. In: Emerging
Technologies for Food Processing, pp. 69–99. Sun, D.-W. Ed., Elsevier Academic Press, London, UK.
Toepfl, S., Mathys, A., Heinz, V., and Knorr, D. (2006). Review: Potential of high hydrostatic pressure and pulsed
electric fields for energy efficient and environmentally friendly food processing. Food Rev Int. 22: 405–425.
Tsong, T.Y., and Astunian, R.D. (1986). Absorption and conversion of electric field energy by membrane
bound ATPases. Bioelectrochem Bioenerg. 15: 457–476.
Urala, N., and Lähteenmäki, L. (2007). Consumers’ changing attitudes towards functional foods. Food Qual
Pref. 18: 1–12.
Van Boekel, M. (2009). Kinetic Modeling of Reactions in Foods. CRC Press/Boca Raton, FL, Taylor &
Francis Group.
Vandresen, S., Quadri, M.G.N., de Souza, J.A.R., and Hotza, D. (2009). Temperature effect on the rheological
behavior of carrot juices. J Food Eng. 92: 269–274. doi:10.1016/j.jfoodeng.2008.11.010.
Vega-Mercado, H., Gongora-Neito, M.M., Barbosa-Canovas, G.V., and Swanson, B.B. (2007). Pulsed electric
fields in food preservations- Chapter 33. In: Handbook of Food Preservation, 2nd ed., pp. 783–813.
LLC, Taylor & Francis Group.
Vidal, O. (2014). First pulsed electric field (PEF) application at industrial scale in beet sugar industry. Sugar Ind.
139(1): 37–39.
Vijayalakshmi, S., Nadanasabhapathi, S., Kumar, R., and Kumar, S.S. (2018). Effect of pH and pulsed electric
field process parameters on the aflatoxin reduction in model system using response surface methodol-
ogy. J. Food Sci. Technol. 1–11.
Vorobiev, E., and Lebovka, N. (2010). Extraction from solid foods and bio suspensions enhanced by electrical
pulsed energy (pulsed electric field, pulsed ohmic heating and high voltage electrical discharges). Food
Eng Rev. 2(2): 95–108.
Wawro, S., Kalinowska, H., and Gruska, R. (2009). The effect of high electric field pulses on activity of sugar
beet o-diphenol oxidase and yeast invertase. J Food Eng. 93: 40–44.
Wiktor, A., Iwaniuk, M., Śledź, M., Nowacka, M., Chudoba, T., and Witrowa-Rajchert, D. (2013). Drying
kinetics of apple tissue treated by pulsed electric field. Dry Technol: An Int J. 31(1): 112–119.
Yeom, H.W., Zhang, Q.H., and Dunne, C.P. (1999). Inactivation of papain by pulsed electric fields in a continu-
ous system. Food Chem. 67: 53–59.
Zhang, Y., Hou, Y., Zhang, Y., Chen, J., Chen, F., Liao, X., and Hu, X. (2012). Reduction of diazinon and dimeth-
oate in apple juice by pulsed electric field treatment. J Sci Food Agric. 92: 743–750. doi:10.1002/jsfa.4636.
Zhao, W., Yang, R., Lu, R., Wang, M., Qian, P., and Yang, W. (2008). Effect of PEF on microbial inactivation
and physical–chemical properties of green tea extracts. LWT-Food Sci Technol. 41: 425–431.
Zhao, W., Yang, R., Zhang, H.Q., Zhang, W., Hua, X., and Tang, Y. (2011). Quantitative and real time detection
of pulsed electric field induced damage on Escherichia coli cells and sublethally injured microbial cells
using flow cytometry in combination with fluorescent techniques. Food Control. 22: 566–573.
Zhijian, S., Xiaojun, L., Yuanyuan, Z., Xiaosong, H., and Chen, F. (2009). Method for reducing organophos-
phorus pesticide in soymilk. China Agricultural University, Beijing, China. Patent No: CN101380041 A.
Zhong, K., Wu, J., Wang, Z., Chen, F., Liao, X., Hu, X., and Zhang, Z. (2007). Inactivation kinetics and sec-
ondary structural change of PEF-treated POD and PPO. Food Chem. 100: 115–123.
Zimmermann, U. (1986). Electrical breakdown, electropermeabilization and electrofusion. Rev Physiol
Biochem Pharmacol. 105: 176–257.
Zimmermann, U., and Benz, R. (1980). Dependence of the electrical breakdown voltage on the charging time
in Valonia utricularis. L Membr Biol. 53: 33–43.
ChaptEr 8
pulse Electric Field processing

of Milk and Milk products
Neelam Upadhyay, C. T. Manoj Kumar, Heena Sharma,

Sanket Borad, and Ashish Kumar Singh
CONtENtS
8.1 Introduction ......................................................................................................................... 129

8.2 Microbial Inactivation Principles........................................................................................ 130
8.3 PEF Processing of Milk and Milk Products ....................................................................... 131
8.4 PEF-Induced Inactivation of Vegetative Microbial Cells ................................................... 132
8.5 PEF Processing of Bacterial Spores Inactivation in Milk .................................................. 133
8.6 PEF Effect on Milk Enzymes ............................................................................................. 134
8.7 PEF Treatment Related Changes on Milk Constituents ..................................................... 136
8.8 Effect of PEF on Physico-chemical and Sensory Properties of Milk and Milk Products .......137
8.9 Synergistic Effect of PEF in Combination with Other Processing Technologies............... 138
8.10 Applications of PEF in Dairy Products .............................................................................. 139
8.11 Conclusion........................................................................................................................... 140
References ...................................................................................................................................... 140
8.1 INtrODUCtION
Milk and milk products constitute an important part of our daily diet. Milk is one of the food
commodities which supplies vital nutrients to people of all age groups. Its nutrition density and
presence of vital nutrients in most appropriate form for digestion, absorption, and assimilation and
is unparalleled. Milk and milk constituents have gained prominence because of increasing scientific
evidence pertaining to their health-promoting and disease-alleviating virtues. Milk has been utilized
since time immemorial as base material for the preparation of a wide array of processed dairy prod-
ucts with different flavour, texture, colour, appearance, and nutritional attributes at domestic and
commercial scale. A large number of unit operations are involved in processing of milk and milk
products with the objective of developing desirable sensory characteristics, longer shelf-life, and
ensured safety at affordable cost. However, how different processing interventions affect the nutri-
tional and therapeutic virtues of milk nutrients is a matter of thorough investigations. Thermal treat-
ments including heating, boiling, pasteurization, thermization, sterilization, and UHT processing are
invariably employed for the manufacture of a wide range of processed dairy products with the prime
129
aim of delivering safe foods with adequate storage stability at ambient temperature. Thermal treat-
ments have been known to denature nutritionally important whey proteins, alter salt balance; initiate
Maillard browning which may be undesirable in certain dairy products, and also cause destruction
of vitamins. Heat-induced changes in milk may lead to reduction in pH, disulphide bridge forma-
tion between β-lactoglobulin and κ-casein, browning, and oxidation of milk lipids. These changes
adversely affect the flavour and colour and may alter certain technological parameters such as heat
stability, gelation, and rennet coagulation properties. Therefore, research investigations pertaining
to processing induced changes on microbiological, physico-chemical, functional, nutritional, and
therapeutic characteristics of liquid milk and various categories of processed dairy products have
increased. Consumer awareness and demand for minimally processed foods have shifted the attention
of researchers towards the application of non-thermal processing interventions like high hydrostatic
pressure (HHP) processing, irradiation, ultrasound, pulsed light technology (PLT), and pulsed elec-
tric field (PEF) as alternatives to thermal processing. However, only a few of them have reached to
the stage of commercialization.
PEF can be defined as a non-thermal method of food preservation that uses short pulses of
electricity for treatment of food and inactivates the microbes present in it. It causes minimal
detrimental effect on food quality attributes and offers consumers high-quality foods. For food
quality attributes, PEF technology is considered superior to traditional thermal processing methods
because it avoids or greatly reduces detrimental changes in the sensory and physico-chemical
properties of foods. Use of electricity for microbial inactivation has been attempted for almost
100 years and initial experimentation targeted electrical energy-induced heat generation and tem-
perature rise for microbial destruction. However, despite the evidence that it can kill microbes, the
technique remained confined to laboratory experimentations. The concept was based on electri-
cal resistance and depended largely on electrical conductivity of food products and later on the
process was termed as “Ohmic Heating.” In the United States a process known as “Electropure”
was patented by Fetterman (1928) and consisted of passing an electric current through milk to
raise its temperature to 70°C leading to lethality of microorganisms. The target microorganisms
were Mycobacterium tuberculosis and Escherichia coli and its utilization for pasteurized milk
at commercial scale continued for few years. Later on, many more attempts were made but all
these approaches were based on heat-induced destruction of microorganisms rather than electri-
cal effect and subsequently terminated due to poor acceptability of such processed products in the
market and consumer concerns about the safety of electrically treated foods. In the early part of
the twenty-first century, there has been revitalized interest among stakeholders in exploitation of
non-thermal processes for delivering “wholesome” fresh-like foods with longer shelf-life and free
from any health hazard. PEF is one such technology where considerable amount of research and
development activities have been initiated, resulting in optimized processing parameters, informa-
tion on PEF-induced changes on quality characteristics, storage stability of processed products,
and equipment for their commercial production. The chapter compiles all relevant information
pertaining to underlying mechanisms of microbial inactivation, optimized process conditions for
enhanced mortality, effect of PEF processing on milk enzymes, nutrients and quality attributes, and
a critical appraisal of experimental designs.
8.2 MICrOBIaL INaCtIVatION prINCIpLES
The PEF technology applies the short pulses of high electric fields with an intensity of
1–100 kV/cm for a period of microseconds to milliseconds. The processing time can be calculated
by multiplying the number of pulses by effective pulse duration. Pulses commonly employed in PEF
PuLse eLeCtriC FieLD ProCessinG oF MiLK AnD MiLK ProDuCts 131
processing could be exponential and square wave. Any basic PEF processing equipment or system
consists of three basic components:
1. A high-voltage pulse generator that supplies electrical energy at the selected voltage. There are a number
of high-voltage power supplies used for PEF systems. Detailed information and performance features
can be referred from the review of Barsotti and Cheftel (1999). The generator is connected to one or
several capacitor(s), which are arranged in parallel manner to store the electrical energy temporarily.
2. A treatment chamber containing two electrodes made up of carbon or metal. The chamber design and
shape of electrode are important fabrication variables with the aim of ensuring uniform field distribu-
tion throughout the chamber and around the electrode. The construction material is also selected to
minimize the electrical shock and incidence of electrochemical interaction between the electrode and
food samples.
3. A control system for monitoring the process parameters including the flow rate, temperature, voltage
gradient, and pulse characteristics. Monitoring the processing conditions and appropriate control
are pre-requisites to ensure the quality of resultant products.
The food product to be treated is placed in treatment chamber. The treatment chamber has two
electrodes which are connected together with a non-conductive material to avoid electrical flow
from one end to other. The distance between these two electrodes is referred to as the ‘treatment
gap’. A high-voltage pulse generator generates pulses of short duration which are then transferred
to electrodes. Electrodes conduct these high-intensity short pulses on food material. As food con-
tains ions and charged molecules, electrical current passes through it. This results in destruction of
microorganisms. The treated product is aseptically packed and stored under refrigeration. In case
of milk, it may serve as electrolyte, the potential difference between the electrodes permits the
movement of charged molecules and electrolysis may occur. Charged particles including minerals
ions, proteins, and living microorganisms have net electrical charge and can migrate during the
application of successive pulses. The deposition of charged particles over the electrodes may lead
to electric field distortion owing to variation in electrical conductivity and electrolysis. The total
energy consumption necessary for high-intensity pulsed electric field (HIPEF) is reported to vary
in the range of 107–201 kJ/I, which is substantially lower than 300 kJ/I of energy used for heating
in HTST milk pasteurization process (Sobrino-López and Martín-Belloso, 2010). Milk and milk
products have been investigated for evaluating the efficacy of PEF interventions in microbial inacti-
vation, prolongation of shelf-life, and other possible applications which are reviewed and discussed.
8.3 pEF prOCESSING OF MILK aND MILK prODUCtS
Milk is a complex mixture of protein, carbohydrate, fat, water, vitamins, minerals, and other
nutrients. These molecules are dispersed in milk in the form of a colloidal suspension, emulsion, or a
true solution. Nutritional superiority of milk and milk products is gaining significance with emerging
evidences suggesting the presence of the bioactives present in it. However, higher moisture content,
neutral pH, and richness of nutrients render it a highly perishable commodity. The exposure of milk
to unhygienic conditions after milking leads to contamination by various emerging pathogens viz.
Escherichia coli, Pseudomonas, Salmonella, Listeria, Staphylococcus, etc. Besides this plethora
of pathogenic microorganisms, some of the spoilage microbes (majorly pschychrotrophic) also find
their way into milk. Thus, thermal processing methods such as pasteurization and sterilization are
used to keep milk safe for consumption. However, these processes result in loss of its natural flavour
and nutritional value, thus posing a challenge for the dairy industry to keep milk safe from contami-
nants without affecting its freshness and nutritional quality. Presently, PEF is successfully applied in
fruit and vegetable juices, milk, yoghurt drink, liquid egg, and other liquid products. The studies are
also conducted on the effect of PEF on nutrients, enzymes, and their functionality. These and other
applications of PEF are discussed in detail in later sections.
8.4 pEF-INDUCED INaCtIVatION OF VEGEtatIVE MICrOBIaL CELLS
PEF treatment exerts microcidal effect by two different ways, often referred to as “electrical
breakdown” followed by “electroporation,” which was proposed by Zimmerman (1986). Electrical
breakdown of cell membrane leads to formation of hydrophilic pores in it as well as forced opening
of protein channels that results in increase in permeability of cell membrane and loss of the semi
or selective permeable nature of cell membrane. According to Zimmerman theory, also known as
electrochemical model, the cell membrane acts as capacitor with low dielectric constant. On both
sides of the membrane, the free charges are present but not inside the phospholipid bilayer that cre-
ates a transmembrane. When the applied electric field strength exceeds the critical value, there will
be formation of reversible or irreversible pores that have the potential of about 10 V. On applying
electric field, the charged particles/channels in membrane align themselves around the membrane
resulting in thinning of membrane.
Barsotti and Cheftel (1999) have provided a detailed mechanism of microbial cell destruction
by applying PEF. Earlier, microbial destruction using PEF technique was considered as first-order
reaction kinetics, but it is only applicable for microbial inactivation of lower degree where experi-
ments were pre-formed in batch-type systems. Recent investigations have indicated tailing during
continuous PEF processing and conditions that result in higher microbial destruction. The inacti-
vation of microorganisms is a complex phenomenon which depends on several factors including
processing factors and product factors. The processing variables like electric field intensity, treat-
ment time, pulse shape, and level of applied energy have a marked effect on microbial destruction
and product quality. Practically, electric field strength should be kept as low as possible for the
effectiveness of treatment; otherwise, high-intensity electrical fields may cause dielectric break-
down of fluids and are conducive to arcing and undesirable reactions. Square waves are considered
superior to exponentially decaying pulses as they decrease linearly from a maximum at peak field
to a minimum rapidly. Square wave pulses are more energy efficient and lethal as microbes are
exposed to higher voltage for longer period of time. Microbial destruction increases in linear man-
ner with processing time. Approximately, 10 μs period is required to establish a transmembrane
potential. After pore formation, the electric field must sustain for 1–5 μs for the pore expansion
to develop permanent damage in microbial cells. However, longer duration would promote elec-
trolysis and electrodeposition at the electrode surface. Product variables like initial microbial load,
type of microorganisms, growth phases, temperature, viscosity, pH conditions, and composition
of the food also impact microbial inactivation. Higher initial microbial load would result in lower
inactivation. Microbial cells in their logarithmic phase are more sensitive to PEF treatment. PEF
treatment is more effective for the destruction of vegetative cells of gram-negative bacteria in com-
parison to the gram-positive bacteria. This is because of the presence of thicker and stouter cell
wall in case of gram-positive bacteria due to the cross-linking of peptidoglycan. Also, lower field
strength has more influence on the larger cells due to the presence of larger surface area available
to be in contact with PEF in comparison to the smaller cells (Ben Ammar et al., 2011). Microbial
inactivation of similar species too varies in different foods. Microbial inactivation also depends on
the conductivity of the medium and microorganisms are more susceptible under lower conductiv-
ity. Effect of pH of the food on microbial inactivation is inconclusive but there exist some reports
indicating that lower pH enhances lethality. This is because the lower pH of the food would affect
the homeostasis of the cells. Low water activity has also been observed to increase the resistance to
PEF-induced inactivation. The reported studies have compared the effect of PEF in buffer medium,
model foods inoculated with microorganisms (reconstituted and/or sterilized skim milk), and real
foods (fresh milk). Sepulveda et al. (2009) suggested a synergistic interaction of PEF and mild heat
treatment for extending the shelf life of milk. The shelf life of whole milk was reported to extend by
a minimum of 24 days after the application of five pulses with 35 kV/cm peak electric field strength
and 2.3 μs pulse width applied to milk at 65°C for less than 10 s.
In model foods, the pathogenic microorganisms such as E. coli, L. monocytogenes, S. aureus,
Salmonella, etc. are inoculated into sterilized skim milk or other dairy products followed by pro-
cessing of milk by PEF to achieve required level of inactivation. Michalac et al. (2003) employed the
continuous PEF bench-scale system which was set to deliver 35 kV/cm field strength with 64 pulses
of bipolar square wave for 188 μs to process raw skim milk and ultra-high-temperature (UHT) skim
milk inoculated with Pseudomonas fluorescens, Lactococcus lactis, and Bacillus cereus at a flow
rate of 1 mL/s. The study achieved 0.3–3 log reduction in all the inoculated microorganisms. Shin
et al. (2007) reported 8-log reduction for E. coli and P. fluorescens and 3-log reduction for Bacillus
stearothermophilus in UHT-treated full-fat milk when exposed to 30–60 kV/cm square wave PEF
with 1 μs pulse width and 26–210 μs treatment time in a continuous PEF treatment system. The effect
of high-intensity PEF processing of infant formula milk inoculated with Enterobacter sakazakii
with varied electric field and treatment time of 10–40 kV/cm and 60–3895 μs (2.5 μs pulse width),
respectively, has been studied by Pina-Perez et al. (2007). The authors have reported a reduction
of inoculated microorganisms by 1.2 log units after exposure to PEF for 360 μs at an intensity of
40 kV/cm. Further, Pina-Perez et al. (2009a) studied processing of infant milk formula inoculated
with Cronobacter sakazakii subsp. sakazakii by using high-intensity PEF with field strength varying
from 15–35 kV/cm at different time and energy levels followed by 24 hours refrigerated storage (8°C)
and the workers reported maximum inhibition of 2.30 log cycles at 15 kV/cm with 3000 μs treatment
time. Pseudomonas species are the major psychrotrophic spoilage micro-organism that affect the
shelf-life of refrigerated milk, stored pasteurized milk and long-life milk (UHT). These bacteria can
grow at lower temperatures and occur widely dispersed in milk production and manufacturing envi-
ronment. Growth of spoilage bacteria, especially psychrotrophic, usually present as post-processing
contaminants is becoming an alarming food safety issue. Several food-borne illnesses are reported
every year because of growth of pathogenic psychrotrophs. Craven et al. (2008) reported that PEF
treatment has the potential to enhance the shelf-life of fresh milk. The researchers carried out an
investigation wherein Pseudomonas isolates were added at levels of 103 and 105 CFU mL−1 into ster-
ile milk (UHT). The results of the research indicated that application of heat without PEF treatment
resulted in only 0.2 logs inactivation of Pseudomonas, while PEF treatment (55°C with 31 kV cm−1)
caused >5 logs reduction. At a level of 103 mL−1 in milk, PEF treatment caused an increase in shelf
life of at least 8 days at 4°C, with total shelf life of at least 13 days. At a level of 105 mL−1 in milk,
PEF treatment increased total shelf life of milk to 11 days. In another study conducted by Guerrero-
Beltrán et al. (2010), the researchers reported around 4.3 log cycle reduction of Listeria innocua on
treating milk with 20, 15, 12.5, 10, and 3 pulses at respectively 3, 15, 23, 33, and 53°C and 40 kV/cm.
The authors suggested that above 53°C, the inactivation of bacteria was dependent less on treatment
time and more on the electric field intensity and treatment temperature.
8.5 pEF prOCESSING OF BaCtErIaL SpOrES INaCtIVatION IN MILK
Spores inactivation remains a challenge in PEF as well as they may cause safety threat. Like
other processing technologies, spore germination followed by PEF treatment was observed to be
effective. Spore inactivation was suggested by using a single or combination of pre-treatments
to induce germination such as mild heating or lysozyme treatment followed by PEF treatment to
inactivate the vegetative cells. Lysozyme may dissolve the outer impervious layer of spores and
render them susceptible for pulse treatment. Bermúdez-Aguirre et al. (2012) reported that Bacillus
cereus spores showed only 1.5 log cycles of inactivation in skim and whole milk after PEF pro-
cessing using 20 pulses (2.5 μs width) at field strength of 30–40 kV/cm at 45°C. The results of
the study were interesting as Bacillus spores showed resistance and germinated soon after PEF
processing, even at temperatures as high as 45°C and 55°C; however, best results were obtained
at 65°C and 75°C. In another study, the authors suggested that Bacillus cereus spores are highly
resistant to high-temperature processing and have a D value of 2 min at 95°C. These spores exhib-
ited resistance over applied electric field strength and germinate, possibly due to stimulation by
electric field, and required severe stress to get inactivated (Byrne et al., 2006). A study conducted
by Pina-Perez et al. (2009b) revealed 3-log reduction of Bacillus cereus vegetative cells inoculated
in skim milk and liquid whole egg (LWE)-skim milk beverage after treating with PEF at 40 kV/cm,
360 μs, and 20°C. Dutreux et al. (2000) reported that application of 63 pulses (2.5 μs width) at an
intensity of 41 kV/cm could reduce E. coli and Listeria innocua by 4 log10 counts. On the contrary,
Bermúdez-Aguirre et al. (2012) reported that application of PEF at higher than 10 pulses at an
intensity of 35 kV/cm at 50°C can increase the temperature above 80°C allowing the milk to boil
inside the treatment chamber and terminating the process because of electric arcing. Salvia-Trujillo
et al. (2011) reported that 1800 μs of PEF treatment at electric field of 35 kV/cm is sufficient to inac-
tivate L. innocua by 5 log units in fruit juice, skim milk, or whole milk. Fleischman et al. (2004)
reported that L. monocytogenes inoculated in skim milk gel showed 1-log reduction when treated
with PEF at 30 kV/cm for 10 pulse. The inactivation remained static even when pulses increased to
50 at electric field strength of 20 kV/cm. PEF at 30 and 40 kV/cm with 50 pulses (square wave with
1 μs width) is reported to reduce the L. innocua by 1.1 and 3.3 log cycles, respectively, in low-fat
UHT milk (Noci et al., 2009). L. innocua inoculated in milk-based smoothie was reduced by 2.7 log
units when exposed to PEF at an electric field of 34 kV/cm with pulse width of 32 μs (Palgan et al.,
2012). Evrendilek et al. (2004) delineated that Staphylococcus aureus inoculated in skim milk were
reduced by 3 log units when exposed to PEF at an intensity of 3.5 kV/mm with treatment time of
460 μs. However, a maximum inactivation of 4.5 log units of Staphylococcus aureus was obtained
by applying 150 bipolar pulses of 8 μs each at 35 kV/cm (Sobrino-López et al., 2006). In this study,
the investigators used bipolar pulses as these are more effective than monopolar. They also observed
that increase in electric field intensity, pulse number, and pulse width resulted in a drop in the sur-
vival fraction of Staphylococcus aureus. Shorter pulses produced greater inactivation than longer
pulses. Rowan et al. (2001) reported that PEF at an intensity of 30 kV/cm applied with 2500 pulses
at 50°C reduced Mycobacterium tuberculosis counts by 5.9 log units when inoculated in cow milk.
8.6 pEF EFFECt ON MILK ENZYMES
Catalytic activity of enzymes relies on the native configuration of their active sites and the con-
figuration of surrounding proteins. The inactivation of enzymes depends on type of enzymes and
conditions adopted during processing. The mechanism of inactivation is speculated to be due to
unfolding, denaturation, and breakdown of covalent bonds and oxidation–reduction reactions in
the protein structure caused by intense electric field (Barsotti and Cheftel, 1999). Inactivation of
enzymes by PEF has been reported by several researchers, but the studies have provided contradic-
tory results. PEF treatment is generally adopted for the purpose of inactivation of microorganisms.
Several researchers have indicated that enzymes are more resistant to PEF and the main variables
that affect the enzyme inactivation are field strength, pulse duration, pulse shape, and number of
pulses (Ho et al. 1997; Vega-Mercado et al., 1997, 2001; Perez and Pilosof, 2004). PEF processing has
been reported to affect secondary and tertiary structures of enzymes and the same has been recorded
by several researchers (Yeom et al., 1999; Zhong et al., 2007). Milk contains several endogenous
enzymes such as phosphatase, lactoperoxidase (LPO), lactoferrin (LF), lysozyme, plasmin, and
exogenous enzymes produced by microorganisms such as proteases, lipases, etc. The presence of
enzymes may create undesirable changes leading to quality defects, so inactivation of these enzymes
is required. Generally, during pasteurization, most of the indigenous enzymes get inactivated, more
importantly alkaline phosphatase (ALP) enzyme, which is used as an indicator of pasteurization
efficiency. However, exogenous enzymes are thermal resistant and require sterilization for their inac-
tivation. Grahl and Markl (1996) observed only a slight inactivation of native ALP (about 5%) after
subjecting raw milk samples to 20 pulses of 21.5 kV/cm with 400 kJ/L energy input (45°C–50°C).
The effect of processing of raw skim milk by PEF at an intensity of 25–37 kV/cm for 200 pulses
per second (2-μs pulse width) at treatment temperature of 15°C and 60°C on ALP was studied by
Shamsi et al. (2008). The investigation concluded 24%–42% inactivation of ALP activity. Zhao and
Yang (2008) studied the effect of PEF processing at different electric field strengths, treatment times,
electrical conductivity, and enzyme concentrations on inactivation of lysozyme enzyme in phosphate
buffer. They reported that relative residual activity (RRA) of lysozyme was decreased with increase
in treatment time at constant electric field intensity. Also, RRA decreased with increase in electric
field strength and maximum reduction in activity of lysozyme (38.1%) was obtained at 35 kV/cm for
1200 μs, which might be due to unfolding of tertiary structures after PEF processing. In addition to
this, more tyrosine residues were buried inside the protein, accompanied by the exposure of more
tryptophan residues after the treatment in protein. Loss of inactivation of lysozyme by PEF was
closely related to the loss of α-helix of secondary structure. However, it was found that the electrical
conductivity is important factor for inactivation of lysozyme; as electrical conductivity increased, the
inactivation rate was decreased. It was speculated that the higher electrical conductivity decreased
the resistance of the medium, and the lower resistance could lead to a negative reflection of the
pulse shape and a lower efficiency of treatment as the system was under loaded. It was also observed
that shape of the pulse became irregular with increase in the electrical conductivity. Further, lyso-
zyme was found more stable and it was suggested that, at higher concentration, lysozyme might have
been stabilized by their tertiary conformation and remained as macro ions to stabilize effectively
the native conformation. However, the study conducted by Yang et al. (2004) revealed no signifi-
cant changes in lysozyme activity after processing with PEF at electric field intensity varying from
0–38 kV/cm for 126 μs treatment time. The reason for no variation in the activity was speculated to
be insufficient treatment time. The effect of different types (batch and continuous flow) of HIPEF
producing equipment on activity of lipase enzyme from P. fluorescens in simulated milk ultrafiltrate
(SMUF) was studied by Bendicho et al. (2002). The workers obtained inactivation of up to 62.1% and
13% of activity by using batch (UDL-B) and continuous (WSU-C) equipment, respectively. However,
Bendicho et al. (2005) reported that protease from Bacillus subtilis in SMUF did not show any change
after treatment under batch mode. On the contrary, the investigators found that activity of protease
decreased by 12% after treatment with HIPEF at 2 Hz, 30.5, or 37.3 kV/cm electric field strength for
84 seconds by using continuous system (WSU-C). However, the protease activity increased in milk
under similar high-intensity PEF processing. Vega-Mercado et al. (2001) suggested that inactivation
of protease depends on the type of media used during HIPEF processing. A study conducted by
Jaeger et al. (2010) to differentiate between effect of electric field and temperature (generated during
PEF treatment) on ALP and LPO in milk. The researchers reported that thermal effects are major
reason for the enzyme inactivation than electric field strength. The activity of ALP and LPO reduced
by 15% and 7% at electric field strength of 34 and 38 kV/cm without exceeding the treatment tem-
perature of 38 and 44°C, respectively. Recently, Sharma et al. (2014b) reported that PEF treatments at
20.7–26.2 kV/cm reduced indigenous ALP activity in whole milk similar to the pasteurized samples.
Also, PEF treatment at 26.1 kV/cm for 34 μs reduced the activity of plasmin, xanthine oxidase, and
lipolysable fat by 12%, 32%, and 82%, respectively, as compared to raw milk. It can be concluded
from the mentioned studies that PEF processing has slight effect on enzymes in milk, so it can be
recommended for cheese making. However, it may not be suitable for inactivation of extracellular
enzymes produced by microorganisms which are critical for the quality of sterilized or UHT milk.
In relation to this, Yu et al. (2012) studied the effect of PEF processing of milk on proteolysis of
cheese. The authors reported that PEF-treated milk showed intermediate proteolysis profile between
raw milk and pasteurized milk in terms of peptides and free amino acid concentration. The impact
of PEF on inactivation of certain enzymes was studied by Riener et al. (2009). The research indicated
a reduction of around 14%, 29%, and 37% in, respectively, lipase, ALP, and protease of fresh bovine
milk at 35 kV/cm for 75 μs treatment time. Recently, Sharma et al. (2014b) reported a reduction in the
activity of plasmin and xanthine oxidase by 12% and 32%, respectively, on application of 26.1 kV/cm
PEF treatment for 34 μs in combination with pre-heating (55°C for 24 s) of bovine whole milk. The
lipolytic activity was reported to decrease up to 82% under the test conditions. There was a strong
correlation between milk enzyme inactivation and electric field intensity.
8.7 pEF trEatMENt rELatED ChaNGES ON MILK CONStItUENtS
PEF is considered a potential replacement for age-old thermal processing (particularly

pasteurization). For this, the research community has studied the effect of PEF processing on milk
constituents so as to establish its effect on the nutritional status of milk. Exposure of macromolecules
to high-intensity electrical fields may alter their structure by different ways including movement of
free electrons and charged particles, displacement of bound compounds, and orientation of protein and
water molecules with constant dipole moment (Castro et al., 2001). PEF treatment also alters the molec-
ular interactions leading to generation of new ions or linkages which could alter the properties of milk.
PEF, at lower intensity treatments, has no significant effect on proteins, vitamins, fat, and other
major or minor nutrients of milk. However, PEF processing variables like intensity, type, and num-
ber of pulses; duration of treatment; and temperature influence the degree of protein denaturation
and modification in milk fat. Any alteration in milk proteins has an influence on the functional
characteristics such as aggregation, hydrophobicity, thermal stability, rennetability, gelation, emulsifi-
cation, and foaming. In milk, major changes are obtained in case of micellar casein including decrease
in casein micelle size, hydrodynamic volume, and loss of calcium from colloidal to soluble phase.
β-lactoglobulin solutions (2, 8, and 12% w/w) when subjected to PEF treatment at electric field
intensity of 30 kV/cm with 200 pulses at 1 Hz frequency did not show marked unfolding and aggre-
gation of β-lactoglobulin (Barsotti et al., 2002). It is well known that the protein aggregation pro-
cess begins with partial unfolding of protein molecules followed by exposure of sulfhydryl groups
and subsequent aggregation through intermolecular disulphide interactions. Perez and Pilosof
(2004) studied the impact of PEF (long pulses and high intensity) on the structural modification of
β-lactoglobulin (12.5 kV/cm). The results of the study indicated a partial denaturation to the extent
of 26%–40% on application of up to 10 pulses of milliseconds. The partial denaturation manifested
as an increased gelation rate of the protein due to PEF processing. Sui et al. (2010) studied the effect
of PEF (electric field strength 35 kV/cm, specific energy 40.6 kJ/kg, and treatment time 19.2 μs) at
different treatment temperatures (30°C, 50°C, 60°C, 65°C, and 70°C) on physico-chemical proper-
ties of bovine LF with different iron saturation levels dissolved in SMUF. They reported that LF
was stable in PEF and non-PEF-treated samples processed between 30°C and 60°C. However, a
decrease in the LF content was observed when treatment temperature increased to 65°C–70°C. The
physico-chemical properties of LF were also altered at higher temperature, which was largely due to
concurrent thermal effects during PEF treatments. PEF treatments do not have an effect on retinol,
α-tocopherol, thiamine, or riboflavin content of raw milk (Riener et al., 2009).
Zhao and Yang (2012) studied the effect of PEF processing on ovalbumin (OVA), bovine serum
albumin (BSA), and a mixture of the two proteins (OVA+BSA) in solution. The OVA showed self-
aggregation through disulphide bonds due to the exposure of sulfhydryl groups and intermolecular
disulphide interactions when PEF intensity exceeded 25 kV/cm. However, no protein self-aggrega-
tion was observed in case of BSA under PEF treatments of 20 to 35 kV cm−1, which might be attrib-
uted to the lack of sulfhydryl groups in BSA molecule. The mixture of the two proteins (OVA+BSA)
showed that both OVA and BSA molecules were involved in the protein interaction and aggregation
when PEF intensity exceeded 25 kV/cm. The BSA which was not sensitive to PEF was incorpo-
rated covalently into protein aggregates through disulphide cross-links with the other proteins. Li
et al. (2003) enriched soymilk with hyper-immunized dairy milk protein concentrate with the aim
to increase bovine IgG level when subjected to PEF processing at 41 kV/cm for 54 μs; the results
showed no significant change in bovine IgG activity.
Garcia-Amezquita et al. (2009) reported that PEF processing of cheese-making milk at
36 kV/cm and 42 kV/cm up to 64 pulses did not modify the true mean diameter of milk fat globule
(MFG), but it induced small globules to clump together, causing an apparent increment in the
population of larger MFGs. Ye et al. (2005) suggested that aggregation of MFGs into larger clumps
might be due to the attachment of whey proteins and heat-affected casein micelles on the surface
of MFG. Hemar et al. (2011) reported that PEF processing (electric field intensity, treatment time,
and specific pulse energy input of 45 kV/cm, 20 μs and 202 kJ/L, respectively) of reconstituted
skim milk (total solids 10%) did not show any significant changes in size of casein micelle after
PEF processing.
8.8 EFFECt OF pEF ON phYSICO-ChEMICaL aND SENSOrY

prOpErtIES OF MILK aND MILK prODUCtS
Milk undergoes certain pre-treatments prior to product manufacture that have marked effect on
various quality attributes and safety of processed dairy products. Thermal treatments like heating,
thermization, and pasteurization are invariably carried out primarily to reduce the initial microbial
load, completely eliminate pathogens, and also inactivate certain milk enzymes. However, heating
often impairs the certain techno-functional properties of milk, including heat stability, colour, dena-
turation of whey proteins, bridge formation, and even development of off-flavour. Industry is looking
for certain alternatives where undesirable changes can be minimized without compromising the
safety aspects and PEF processing appears to be a viable alternative.
The effect of PEF processing at different temperature (20°C, 30°C, and 40°C) and electric field
strengths (30.76 to 53.84 kV/cm) on the physico-chemical parameters (pH, electrical conductivity,
density, colour, solids not fat) of skim and whole milk was studied by Bermudez‐Aguirre et al. (2011).
Minor variations in physico-chemical properties of skim and whole milk after PEF processing were
observed, unlike thermal treatment. However, few other researchers have reported no major varia-
tion in electrical conductivity and density of milk on PEF processing. PEF processing (electric field
intensity 45 kV/cm, treatment time 20 μs, and specific pulse energy input 202 kJ/L) of milk concen-
trate (total solids 18%) also did not show any significant changes in viscosity after PEF processing
(Hemar et al., 2011). However, the minor reduction in milk viscosity in PEF-treated milk could be
attributed to modification of hydrodynamic volume of caseins and partly to alteration in salt bal-
ance. However, a slight lowering in SNF content and total solids in milk has been reported. This
could probably due to deposition of SNF over electrodes owing to electrolysis of milk.
Xiang et al. (2011) studied the effect of PEF processing at different electric field intensities (12, 16,
and 20 kV/cm) and number of pulses (10, 20, and 30) on the several physico-chemical properties of
whey protein isolates (WPI). PEF processing of WPI increased the polarity of tryptophan residues
in whey proteins, causing partial denaturation of WPI fractions, and exposure of more hydrophobic
regions under these PEF treatments. However, Sui et al. (2011) found that PEF treatment (electric field
intensity, pulse width, frequency, treatment time, and specific pulsing energy input of, respectively,
35 kV/cm, 2 μs, 100 Hz, 19.2 μs, and 131.9 kJ/L) did not affect any of the physico-chemical or emulsifi-
cation properties of WPI solutions (1% w/w). However, the heat-induced gel strength of WPI decreased
from 461 Pa to 139 and 67 Pa after PEF treatment at 30 kV/cm for 19.2 and 211 μs, respectively, and the
corresponding gelation times increased from 40.8 min to 43.2 and 47.9 min, respectively.
Cheese, a leading dairy product, is of high commercial value all over the globe on account of
being rich in nutrients and availability of wide varieties. Certain varieties of cheese are preferred to
be prepared using raw milk, commonly referred to as raw milk cheese. These cheeses are famous
due to distinctive flavour and texture, not replicable by using pasteurized milk. However, such
cheeses are prone to be associated with cheese-related outbreaks especially when the hygienic milk
production practices are not followed. Thus, the milk used for preparation of cheese is required
to be pasteurized for inactivation of bacteria, but this is accompanied by adversely affecting the
flavour, taste, and nutrients of product (if inactivation is caused by thermal treatment). In this
regard, Sepulveda-Ahumada et al. (2000) applied high-voltage PEF to pasteurize milk used for
cheese making. Textural and sensorial analysis of cheddar cheese prepared from PEF-processed
milk revealed a harder and springier cheese than the one prepared from treated milk, while adhe-
siveness and cohesiveness were not significantly different. Sensory scores showed significant
differences in flavour profile of cheeses, but no significance variation in sensory textural parameters
was observed. Variations in flavour were attributed to release of sulfhydryl compounds from cys-
teine and cystine at elevated temperature generated during high-voltage PEF. A few investigations
also indicated that PEF treatment resulted in attenuation of lactic starters with higher proteolytic
and peptidolytic activity. Yu et al. (2009) compared the coagulation properties of PEF-treated milk,
thermally pasteurized milk, and raw milk. The researchers reported that within the experimental
range (electric field intensity of maximum 30 kV/cm, 120 pulses, and treatment temperature up to
50°C), PEF-treated milk resulted in better rennetability in comparison to thermally pasteurized
milk. Thus, the researchers advocated PEF (electric field strength of ≤30 kV/cm) in combination
with mild thermal treatment (≤50°C) to be potential pasteurization method for the milk aimed for
the production of cheese. Most of the experiments indicate the positive effect of PEF processing on
quality characteristics of processed dairy products, but compromising safety concerns. However,
well-planned experimentation with larger batches is desired to draw a logical conclusion for the
applicability of PEF processing of milk and other dairy products at commercial scale.
8.9 SYNErGIStIC EFFECt OF pEF IN COMBINatION

WIth OthEr prOCESSING tEChNOLOGIES
PEF is a novel non-thermal technology that has shown positive results in inactivation of pathogenic
and spoilage causing microorganisms. However, several factors such as food composition, electrical
conductivity, temperature, type of microorganisms, etc., limit their effectiveness against certain level
of inactivation. Generally, a process which is able to reduce microbial population by >5 log CFU/g
is considered as a disinfection process. Several studies have shown that PEF alone is not effective in
reducing the spoilage and pathogenic microorganisms. Hence, PEF is required to be combined with
other antimicrobial processes or agents to get a desirable level of inactivation. Several studies have
been conducted in this view to have synergistic effect on inactivation of microorganisms, which are
discussed here after.
Sharma et al. (2014a) reported that PEF (electric field strength 18–28 kV/cm for 17–101 μs)
processing of milk at 4°C did not affect the bacterial population of Pseudomonas aeruginosa,
Escherichia coli, Staphylococcus aureus, and Listeria innocua inoculated in milk. However,
when temperature increased from 4°C to 55°C, complete inactivation of all inoculated bacteria
was observed. It could be due to the increase in fluidity of cell membrane at elevated temperature
leading to reduction in their mechanical resistance. P. aeruginosa and E. coli (gram-negative) were
less resistant to PEF process compared to S. aureus and L. innocua (gram-positive). Pina-Perez
et al. (2009c) conducted a study for the purpose of extending the shelf-life and enhancing the safety
of LWE and skim milk synergistically by combining PEF processing (at 40 kV/cm, 360 μs, 20°C)
with addition of 12% of CocoanOX (i.e., antimicrobial cocoa powder). The product with or without
CocoanOX (12%) was inoculated with Bacillus cereus and was stored at 5°C for 15 days after treat-
ment with PEF. Both the treated products showed 3 log cycles reduction of B. cereus, whereas cocoa
supplemented products showed 3.30 log cycle reduction. After 15 days of storage, the antimicrobial
compound lowered the B. cereus counts in the samples supplemented with CocoanOX (12%) by a
4-log cycle reduction, as compared to the untreated samples. Sepulveda et al. (2005) attempted to
develop extended shelf-life milk by combining high-temperature short-term (HTST) pasteurization
with PEF. HTST-pasteurized milk immediately or after 8 days of pasteurization was PEF processed
at peak electric field strength of 35 kV/cm with pulse width of 2.5 μs at 65°C for less than 10 s,
which enhanced the shelf-life of milk to 60 and 78 days, respectively. It indicated that prior heating
of raw milk followed by PEF treatment had an additive effect on microbial destruction. Noci et al.
(2009) processed milk inoculated with L. innocua with thermo-sonication at 400 W power followed
by PEF treatment at peak electric field strength of 40 kV/cm. The study revealed a reduction in
L. innocua counts by 6.8 log cycles. This indicates that PEF combined with thermo-sonication can
be considered a suitable alternate to thermal processing of milk. Pina-Perez et al. (2012) studied the
synergistic effect of combination of cinnamon and PEF on inactivation of Salmonella typhimurium
inoculated in skim milk beverage. The researchers could achieve maximum inactivation of 1.97 log
cycles using a hurdle approach consisting of a combination of PEF treatment at electric field strength
30 kV/cm, treatment time 700 μs, and cinnamon application at 5 percent level. Gallo et al. (2007)
studied the efficiency of the combined treatment of nisin and PEF and observed an antagonistic
effect on L. innocua inoculated in liquid whey protein concentrate (8% solution). This could be due
to change in cell envelope that might have been caused by increase in the cell resistance to nisin
and partly to modification of the medium. However, when nisin was added prior to PEF treatment,
it exhibited an additive and slightly synergistic effect, suggesting that the binding of nisin to the
cell membrane would increase the susceptibility of the microorganism to PEF treatment. PEF pro-
cessing alone at lower temperature may not be considered an effective approach for enhancing the
shelf-life because of limited lethality. However, if it is combined with thermal treatment at modest
temperature, with anti-microbial compounds or other preservation techniques, the effect is more
pronounced. Hence, PEF can constitute as important part of a hurdle system for enhancing the
shelf-life of heat-sensitive products like vitamin-fortified beverages, probiotic dairy drinks, and
viscous health beverages such as smoothies, etc.
8.10 appLICatIONS OF pEF IN DaIrY prODUCtS
The applications of PEF are generally limited to liquid foods, especially the ones that have
lower electrical conductivity and which do not contain bubbles. Milk can thus be a potential product
where PEF can be used effectively. Yeom et al. (2004) observed the effect of mild heat treatment
alone and in combination with PEF treatment on the microbial stability of yoghurt-based products.
The results of the investigation provide some interesting results as the product treated with PEF
showed significantly decreased total viable aerobic bacteria and total yeast and mould counts in com-
parison to when the heat treatment alone was deployed for checking the growth of microorganisms.
A comparative analysis of PEF treatment (15–30 kV/cm 25°C, 45°C and 65°C), HHP treatment
(450–650 MPa at 30C and 50°C), and thermal treatment (60°C–90°C for 1 min) on the activity of
pectin methylesterase and concentration of volatile compounds in orange juice-milk based bever-
age was carried out by Sampedro et al. (2009). Pectin methylesterase spoilage is reported to cause
cloud loss or gelification of the beverage. The results revealed that 90% of the pectin methylester-
ase could be inactivated using PEF treatment (25 kV/cm, 65°C), HHP treatment (650 MPa, 50°C),
and thermal treatment (85°C, 1 min). The researchers extracted twelve volatile compounds using
solid-phase microextraction method and quantification of these was done using GC-MS. A loss of
volatiles was reported following different treatments. However, the minimum loss was observed for
PEF treatment, i.e., 13.7% to 8.3% at 25°C, 5.8% to 21.0% at 45°C, and 11.6% to 30.3% at 65°C. The
authors suggested that pulp-related compounds were more resistant to HHP and PEF, while higher
molecular weight compounds were more resistant to thermal treatments. The authors advocated
the industrial relevance of PEF as the non-thermal technique for the pasteurization of the orange
juice-milk-based beverage and preservation of natural quality of the beverage in terms of retention
of volatile compounds.
A recent work carried out by McAuley et al. (2016) suggested that PEF treatment is good for
maintaining safety without compromising the quality. The authors suggested a similar microbial
quality of thermally treated milk (15 s, at 63°C and 72°C) and PEF-treated milk (30 kV/cm and
22 μs at outlet temperature of 53°C and 63°C). The authors did not report any adverse effect on the
physico-chemical properties of milk.
8.11 CONCLUSION
Consumer demand for fresh-like food products containing native nutrients but safe and free from
any additive and contaminant is increasing day by day. Moreover, the energy auditing in pursuit
of providing processed food with certain novelty at affordable cost also necessitates the search for
alternatives. Among the non-thermal processing technologies, PEF has great potential to meet the
expectations of stakeholders on a commercial scale. Research findings indicate that PEF technology
has potential to inactivate the microorganisms contaminating milk and other dairy products without
adversely affecting the organoleptic, nutritional, or inherent features of the food products. Microbial
inactivation using PEF interventions depends on several processing and product features. Most of
the findings are outcomes of laboratory- or pilot-scale experimentations and their validation in com-
mercial situations is of paramount importance to draw the logical conclusion. Moreover, there is
also need to optimize the PEF processing conditions to achieve the commercial sterility. Checking
of post-processing contamination through appropriate packaging technologies also requires
in-depth investigations. Combination of low intensity thermal treatments, application of natural or
bio-preservatives, and suitable packaging technology with PEF processing has potential for manu-
facturing long-life processed dairy products. No or little changes in composition, physico-chemical,
nutritional, and sensory characteristics of milk and dairy products in PEF-processed foods are added
advantages. The promising applications could be processing and preservation of liquid milk, forti-
fied dairy products, fermented and probiotic dairy drinks, smoothies, and other nutritional beverage
where low temperature processing is of prime significance to improve the quality of finished products.
rEFErENCES
Barsotti, L., and Cheftel, J.C. (1999). Food processing by pulsed electric fields. II. Biological aspects. Food
Rev Int. 15(2): 181–213.
Barsotti, L., Dumay, E., Mu, T.H., Diaz, M.D.F., and Cheftel, J.C. (2002). Effects of high voltage electric
pulses on protein-based food constituents and structures. Trends Food Sci Technol. 12(3–4): 136–144.
Ben Ammar, J., Lanoisellé, J.L., Lebovka, N.I., Van Hecke, E., and Vorobiev, E. (2011). Impact of a pulsed
electric field on damage of plant tissues: Effects of cell size and tissue electrical conductivity. J Food Sci.
76(1): 90–97.
Bendicho, S., Estela, C., Giner, J., Barbosa-Cánovas, G.V., and Martin, O. (2002). Effects of high intensity
pulsed electric field and thermal treatments on a lipase from Pseudomonas fluorescens. J Dairy Sci.
85(1): 19–27.
Bendicho, S., Marsellés-Fontanet, A.R., Barbosa-Cánovas, G.V., and Martín-Belloso, O. (2005). High inten-
sity pulsed electric fields and heat treatments applied to a protease from Bacillus subtilis. A comparison
study of multiple systems. J Food Eng. 69(3): 317–323.
Bermúdez-Aguirre, D., Dunne, C.P., and Barbosa-Cánovas, G.V. (2012). Effect of processing parameters on
inactivation of Bacillus cereus spores in milk using pulsed electric fields. Int Dairy J. 24(1): 13–21.
Bermúdez‐Aguirre, D., Fernández, S., Esquivel, H., Dunne, P.C., and Barbosa‐Cánovas, G.V. (2011). Milk
processed by pulsed electric fields: Evaluation of microbial quality, physicochemical characteristics,
and selected nutrients at different storage conditions. J Food Sci. 76(5): 289–299.
Byrne, B., Dunne, G., and Bolton, D.J. (2006). Thermal inactivation of Bacillus cereus and Clostridium perfrin-
gens vegetative cells and spores in pork luncheon roll. Food Microbiol. 23(8): 803–808.
Castro, A.J., Swanson, B.G., Barbosa-Canovas, G.V., and Zhang, Q.H. (2001). Pulsed electric field modification
of milk alkaline phosphatase activity. In: Pulsed Electric Fields in Food Processing. Fundamental Aspects
and Applications, pp. 65–82. Barbosa-Canovas, G.V., and Zhang, Q.H. Eds., Technomic Publishing
Company Inc., Lancaster, PA.
Craven, H.M., Swiergon, P., Ng, S., Midgely, J., Versteeg, C., Coventry, M.J., and Wan, J. (2008). Evaluation
of pulsed electric field and minimal heat treatments for inactivation of pseudomonads and enhancement
of milk shelf-life. Innov Food Sci Emerg Technol. 9(2): 211–216.
Dutreux, N., Notermans, S., Wijtzes, T., Gongora-Nieto, M.M., Barbosa-Canovas, G.V., and Swanson, B.G.
(2000). Pulsed electric fields inactivation of attached and free-living Escherichia coli and Listeria
innocua under several conditions. Int J Food Microbiol. 54(1–2): 91–98.
Evrendilek, G.A., Zhang, Q.H., and Richter, E.R. (2004). Application of pulsed electric fields to skim milk
inoculated with Staphylococcus aureus. Biosyst Eng. 87(2): 137–144.
Fetterman, J.C. (1928). The electrical conductivity method of processing milk. Agr Eng. 9(4): 107–108.
Fleischman, G.J., Ravishankar, S., and Balasubramaniam, V.M. (2004). The inactivation of Listeria mono-
cytogenes by pulsed electric field (PEF) treatment in a static chamber. Food Microbiol. 21(1): 91–95.
Gallo, L.I., Pilosof, A.M., and Jagus, R.J. (2007). Effect of the sequence of nisin and pulsed electric fields
treatments and mechanisms involved in the inactivation of Listeria innocua in whey. J Food Eng. 79(1):
188–193.
Garcia-Amezquita, L.E., Primo-Mora, A.R., Barbosa-Canovas, G.V., and Sepulveda, D.R. (2009). Effect of
non-thermal technologies on the native size distribution of fat globules in bovine cheese-making milk.
Innov Food Sci Emerg Technol. 10(4): 491–494.
Grahl, T., and Märkl, H. (1996). Killing of microorganisms by pulsed electric fields. Appl Microbiol
Biotechnol. 45(1–2): 148–157.
Guerrero-Beltrán, J.Á., Sepulveda, D.R., Góngora-Nieto, M.M., Swanson, B., and Barbosa-Cánovas, G.V.
(2010). Milk thermization by Pulsed electric fields (PEF) and electrically induced heat. J Food Eng.
100(1): 56–60.
Hemar, Y., Augustin, M.A., Cheng, L.J., Sanguansri, P., Swiergon, P., and Wan, J. (2011). The effect of pulsed
electric field processing on particle size and viscosity of milk and milk concentrates. Milchwissenschaft.
66(2): 126.
Ho, S.Y., Mittal, G.S., and Cross, J.D. (1997). Effects of high field electric pulses on the activity of selected
enzymes. J. Food Engineering. 31: 69–85.
Jaeger, H., Meneses, N., Moritz, J., and Knorr, D. (2010). Model for the differentiation of temperature and
electric field effects during thermal assisted PEF processing. J Food Eng. 100(1): 109–118.
Li, S.Q., Zhang, Q.H., Lee, Y.Z., and Pham, T.V. (2003). Effects of pulsed electric fields and thermal processing
on the stability of bovine immunoglobulin G (IgG) in enriched soymilk. J Food Sci. 68(4): 1201–1207.
McAuley, C.M., Singh, T.K., Haro-Maza, J.F., Williams, R., and Buckow, R. (2016). Microbiological and
physicochemical stability of raw, pasteurised or pulsed electric field-treated milk. Innov Food Sci
Michalac, S., Alvarez, V.T.J.I., Ji, T., and Zhang, Q.H. (2003). Inactivation of selected microorganisms and
properties of pulsed electric field processed milk. J Food Process Preserv. 27(2): 137–151.
Noci, F., Walkling-Ribeiro, M., Cronin, D.A., Morgan, D.J., and Lyng, J.G. (2009). Effect of thermosonication,
pulsed electric field and their combination on inactivation of Listeria innocua in milk. Int Dairy J. 19(1):
30–35.
Palgan, I., Muñoz, A., Noci, F., Whyte, P., Morgan, D.J., Cronin, D.A., and Lyng, J.G. (2012). Effectiveness
of combined pulsed electric field (PEF) and manothermosonication (MTS) for the control of Listeria
innocua in a smoothie type beverage. Food Cont. 25(2): 621–625.
Perez, O.E., and Pilosof, A.M. (2004). Pulsed electric fields effects on the molecular structure and gelation of
β-lactoglobulin concentrate and egg white. Food Res Int. 37(1): 102–110.
Pina-Perez, M.C., Martínez-López, A., and Rodrigo, D. (2012). Cinnamon antimicrobial effect against
Salmonella typhimurium cells treated by pulsed electric fields (PEF) in pasteurized skim milk beverage.
Food Res Int. 48(2): 777–783.
Pina-Perez, M.C., Rodrigo, D., and López, A.M. (2009a). Sub-lethal damage in Cronobacter sakazakii subsp.
sakazakii cells after different pulsed electric field treatments in infant formula milk. Food Cont. 20(12):
1145–1150.
Pina-Perez, M.C., Silva-Angulo, A.B., Muguerza-Marquinez, B., Aliaga, D.R., and López, A.M. (2009b).
Synergistic effect of high hydrostatic pressure and natural antimicrobials on inactivation kinetics of
Bacillus cereus in a liquid whole egg and skim milk mixed beverage. Foodborne Pathog Dis. 6(6):
649–656.
Pina-Perez, M.C., Silva-Angulo, A.B., Rodrigo, D., and Martínez-López, A. (2009c). Synergistic effect of
pulsed electric fields and CocoanOX 12% on the inactivation kinetics of Bacillus cereus in a mixed
beverage of liquid whole egg and skim milk. Int J Food Microbiol. 130(3): 196–204.
Pina-Perez, M.P., Aliaga, D.R., Bernat, C.F., Enguidanos, M.R., and López, A.M. (2007). Inactivation of
Enterobacter sakazakii by pulsed electric field in buffered peptone water and infant formula milk. Int
Dairy J. 17(12): 1441–1449.
Riener, J., Noci, F., Cronin, D.A., Morgan, D.J., and Lyng, J.G. (2009). Effect of high intensity pulsed electric
fields on enzymes and vitamins in bovine raw milk. Int J Dairy Technol. 62(1): 1–6.
Rowan, N.J., MacGregor, S.J., Anderson, J.G., Cameron, D., and Farish, O. (2001). Inactivation of
Mycobacterium paratuberculosis by pulsed electric fields. Appl Environ Microbiol. 67(6): 2833–2836.
Salvia-Trujillo, L., Morales-de la Pena, M., Rojas-Grau, M.A., and Martin-Belloso, O. (2011). Microbial and
enzymatic stability of fruit juice-milk beverages treated by high intensity pulsed electric fields or heat
during refrigerated storage. Food Control 22(10): 1639–1646.
Sampedro, F., Geveke, D.J., Fan, X., and Zhang, H.Q. (2009). Effect of PEF, HHP and thermal treatment on
PME inactivation and volatile compounds concentration of an orange juice–milk based beverage. Innov
Food Sci Emerg Technol. 10(4): 463–469.
Sepulveda-Ahumada, D.R., Ortega-Rivas, E., and Barbosa-Cánovas, G.V. (2000). Quality aspects of ched-
dar cheese obtained with milk pasteurized by pulsed electric fields. Food Bioproduct Process. 78(2):
65–71.
Sepulveda, D.R., Góngora-Nieto, M.M., Guerrero, J.A., and Barbosa-Cánovas, G.V. (2005). Production of
extended-shelf life milk by processing pasteurized milk with pulsed electric fields. J Food Eng. 67(1):
81–86.
Sepulveda, D.R., Góngora-Nieto, M.M., Guerrero, J.A., and Barbosa-Cánovas, G.V. (2009). Shelf life of
whole milk processed by pulsed electric fields in combination with PEF-generated heat. LWT-Food Sci
Technol. 42(3): 735–739.
Shamsi, K., Versteeg, C., Sherkat, F., and Wan, J. (2008). Alkaline phosphatase and microbial inactivation by
pulsed electric field in bovine milk. Innov Food Sci Emerg Technol. 9(2): 217–223.
Sharma, P., Bremer, P., Oey, I., and Everett, D.W. (2014a). Bacterial inactivation in whole milk using pulsed
electric field processing. Int Dairy J. 35(1): 49–56.
Sharma, P., Oey, I., Bremer, P., and Everett, D.W. (2014b). Reduction of bacterial counts and inactivation of
enzymes in bovine whole milk using pulsed electric fields. Int Dairy J. 39(1): 146–156.
Shin, J.K., Jung, K.J., Pyun, Y.R., and Chung, M.S. (2007). Application of pulsed electric fields with square
wave pulse to milk inoculated with E. coli, P. fluorescens, and B. stearothermophilus. Food Sci Biotech.
16(6): 1082–1084.
Sobrino-López, A., and Martín-Belloso, O. (2010). Potential of high-intensity pulsed electric field technology
for milk processing. Food Eng Rev. 2(1): 17–27.
Sobrino-López, A., Raybaudi-Massilia, R., and Martín-Belloso, O. (2006). High-intensity pulsed electric field
variables affecting Staphylococcus aureus inoculated in milk. J Dairy Sci. 89(10): 3739–3748.
Sui, Q., Roginski, H., Williams, R.P., Versteeg, C., and Wan, J. (2010). Effect of pulsed electric field and ther-
mal treatment on the physicochemical properties of lactoferrin with different iron saturation levels. Int
Dairy J. 20(10): 707–714.
Sui, Q., Roginski, H., Williams, R.P., Versteeg, C., and Wan, J. (2011). Effect of pulsed electric field and
thermal treatment on the physicochemical and functional properties of whey protein isolate. Int Dairy J.
21(4): 206–213.
Vega-Mercado, H., Martin-Belloso, O., Qin, B.L., Chang, F.J., Góngora-Nieto, M.M., Barbosa-Cánovas,
G.V., and Swanson, B.G. (1997). Non-thermal food preservation: Pulsed electric fields. Trends Food Sci
Technol. 8(5): 151–157.
Vega-Mercado, H., Powers, J.R., Martı ́n-Belloso, O., Luedecke, L., Barbosa-Cánovas, G.V., and Swanson, B.G.
(2001). Change in susceptibility of proteins to proteolysis and the inactivation of an extracellular protease
from Pseudomonas fluorescens M3/6 when exposed to pulsed electric fields. In: Pulsed Electric Fields
in Food Processing. Fundamental Aspects and Applications, pp. 105–120. Barbosa-Cánovas, G.V., and
Zhang, Q.H. Eds., Technomic Publishing Company Inc., Lancaster, PA.
Xiang, B.Y., Ngadi, M.O., Ochoa-Martinez, L.A., and Simpson, M.V. (2011). Pulsed electric field-induced
structural modification of whey protein isolate. Food Bioprocess Technol. 4(8): 1341–1348.
Yang, R.J., Li, S.Q., and Zhang, Q.H. (2004). Effects of pulsed electric fields on the activity of enzymes in
aqueous solution. J Food Sci. 69(4): 241–248.
Ye, A., Singh, H., Taylor, M., and Anema, S. (2005). Disruption of fat globules during concentration of whole
milk in a pilot scale multiple‐effect evaporator. Int J Dairy Technol. 58(3): 143–149.
Yeom, H.W., Evrendilek, G.A., Jin, Z.T., and Zhang, Q.H. (2004). Processing of yogurt‐based products with
pulsed electric fields: Microbial, sensory and physical evaluations. J Food Process Preserv. 28(3):
161–178.
Yeom, H.W., Zhang, Q.H., and Dunne, C.P. (1999). Inactivation of papain by pulsed electric fields in a continu-
ous system. Food Chem. 67(1): 53–59.
Yu, L.J., Ngadi, M., and Raghavan, G.S.V. (2009). Effect of temperature and pulsed electric field treatment on
rennet coagulation properties of milk. J Food Eng. 95(1): 115–118.
Yu, L.J., Ngadi, M., and Raghavan, V. (2012). Proteolysis of cheese slurry made from pulsed electric field-
treated milk. Food Bioprocess Technol. 5(1): 47–54.
Zhao, W., and Yang, R. (2008). The effect of pulsed electric fields on the inactivation and structure of lysozyme.
Food Chem. 110(2): 334–343.
Zhao, W., and Yang, R. (2012). Pulsed electric field induced aggregation of food proteins: Ovalbumin and
bovine serum albumin. Food Bioprocess Technol. 5(5): 1706–1714.
Zhong, K., Wu, J., Wang, Z., Chen, F., Liao, X., Hu, X., and Zhang, Z. (2007). Inactivation kinetics and
secondary structural change of PEF-treated POD and PPO. Food Chem. 100(1): 115–123.
Zimmermann, U. (1986). Electrical breakdown, electropermeabilization and electrofusion. Rev Physiol
Biochem Pharm. 105: 175–256.
ChaptEr 9
application of Ultrasonic in Food processing
Pradeep Singh Negi and Navin Kumar Rastogi
CONtENtS
9.1 Introduction .......................................................................................................................... 145

9.2 Ultrasonic Equipment, Engineering Aspects, and Principles .............................................. 147
9.3 Applications of Ultrasonics in Food Processing .................................................................. 149
9.3.1 Ultrasonic for Microbial Inactivation ....................................................................... 150
9.3.2 Ultrasonics in Combination with Other Treatments for Microbial Inactivation ...... 150
9.3.3 Ultrasonic-Assisted Enzyme Inactivation ................................................................ 153
9.3.4 Ultrasonic-Assisted Extraction of Biomolecules ...................................................... 155
9.3.5 Ultrasonic-Assisted Drying ...................................................................................... 155
9.3.6 Ultrasonic-Assisted Freezing.................................................................................... 158
9.3.7 Ultrasonic-Assisted Improvement in Animal Product Quality ................................ 159
9.4 Miscellaneous Applications of Ultrasonics in Food Industry .............................................. 160
9.5 Conclusion ............................................................................................................................ 162
References ...................................................................................................................................... 163
9.1 INtrODUCtION
Ultrasonics, a non-thermal processing technique, is one of the fast, versatile, and promising
non-destructive green technologies used in the food industry in the last several years in various
areas of food processing, i.e., blanching, freezing, degassing, extraction, drying, filtration, emul-
sification, sterilization, cutting, and other applications (Figure 9.1). It is being applied as an effec-
tive preservation tool for many categories of food such as fruits and vegetables, cereals and their
products, dairy products, honey, proteins, and meats. Its applications include microbial and enzyme
inactivation and water treatment, to name a few. Ultrasonic waves travel through a medium similar
to sound waves in a series of compressions and rarefactions, and at a sufficiently high power, the
rarefaction exceeds the attractive forces between the molecules in a liquid phase, which subse-
quently leads to the formation of cavitation bubbles. Each bubble affects the localized field experi-
enced by neighbouring bubbles, which causes the cavitation bubble to become unstable and collapse
thereby releasing energy for many chemical and mechanical effects. The cavitation collapse in
aqueous medium generates shear forces that can produce mechanical effects. The collapse of a
cavitation bubble on or near a surface is asymmetrical because the surface provides resistance to
liquid flow resulting in an inrush of liquid (mostly from the side of the bubble), which leads to the
145
Figure 9.1 Applications of ultrasonic in food processing.
Figure 9.2 bubble cavitation formation by ultrasound treatment. (From Kadam et al., Trends Food Sci.
Technol., 46, 60–67, 2015.)
formation of a powerful liquid jet targeted at the surface (Figure 9.2). This effect can also increase
mass transfer to the surface making it equivalent to a high-pressure jetting, which makes ultrasound
suitable for cleaning (Rastogi, 2011).
The physical, mechanical, or chemical effects of high-power ultrasound (20 and 100 kHz) are
capable of altering material properties through the generation of pressure, shear, and temperature
gradient in the medium. Generally, ultrasound equipment uses frequencies from 20 kHz to 10 MHz
(Figure 9.3). Higher-power ultrasound or power ultrasound at lower frequencies (20–100 kHz) has
the ability to cause cavitations (implosion of gas bubbles) with sound intensities of 10–1000 W/cm2
APPLiCAtion oF uLtrAsoniC in FooD ProCessinG 147
Hz 1
Infrasound 10 16 Hz
100
Audiblesound
kHz 1
20 kHz
10 18 kHz
Power ultrasound
100 100 kHz

Extended for
Ultrasound
special applications
MHz 1
5 MHz
Diagnostic purpose 1 MHz
10
10 MHz
100
GHz 1
Figure 9.3 Frequency ranges of ultrasound used in food industry.
(McClements, 1995; Feng and Yang, 2005), and it has been used for the functional modification of
proteins derived from different food sources such as dairy, animal, cereal, legume, tuber, and fruit
(O’Sullivan et al., 2017).
9.2 ULtraSONIC EQUIpMENt, ENGINEErING aSpECtS, aND prINCIpLES
Ultrasound is a form of vibrational energy in the lower frequency range (20–100 kHz with a sound
intensity of 10–1000 W/cm2) that is also referred to as “power ultrasound.” It has the ability to cause
cavitation (implosion of gas bubbles), which is used in food processing to inactivate microorganisms
(Feng and Yang, 2005). Low-intensity ultrasound provides information about physico-chemical prop-
erties, while high-intensity ultrasound is used to alter, either physically or chemically, the properties
of foods, e.g., to generate emulsions, disrupt cells, promote chemical reactions, inhibit enzymes,
tenderize meat, and modify crystallization processes (McClements, 1995).
The ultrasonic transducers convert electrical or mechanical energy to sound energy. There are
three types of ultrasonic transducers in common usage: liquid-driven transducers, magnetostric-
tive transducers, and piezoelectric transducers (Mason, 1998), with piezoelectric being the most
common. For ultrasonic baths, power is often low in order to avoid damage to the tank walls and
the power density is low due to large volume or processing liquid. When high-power ultrasound
propagates in a liquid, cavitation bubbles will be generated due to pressure changes. These micro
bubbles will collapse violently in the succeeding compression cycles of a propagated sonic wave.
These results in regions of high localized temperatures up to 5000 K and pressure of up to 50 MPa,
resulting in high shearing effects and a localized sterilization effect (Piyasena et al., 2003).
Cavitation intensity can be estimated by measuring hydrogen peroxide (H 2O2) formation
in distilled water during sonication following a catalyzed colorimetric procedure (Mead et al.,
1976). However, the determination of H 2O2 generation during an ultrasound treatment in a food
system is complex due to the presence of food components including ions and other colloidal
components. Tsukamoto et al. (2004) reported that the measurement of ultrasound amplitude
is an indication of the ultrasonic cavitation and is also a reliable method for indication of the
ultrasound power.
The two most common pieces of laboratory equipment used for processing liquids are an ultra-
sonic cleaning bath, which is inexpensive and is commonly used to sonicate liquid samples in
vessels immersed in the bath (Figure 9.4a), or the more powerful probe system, which introduces
vibrations directly into the sample (Figure 9.4b) (Mason, 1999). The sonication treatment and
the cavitation activity in a treatment chamber may vary for the same ultrasound intensity if the
sample volume and probe location change. The results from successful small-scale experiments
can be adapted for large-scale work providing that information is available on power input and
volume treated.
Several scale-up designs are available for food processing which can be broadly divided into
batch and flow systems. A general review of large-scale processors has been published by Mason
and Peters (2002). Batch systems will generally be based upon the ultrasonic cleaning bath using
the whole bath as the reactor. It can be found in cleaning and decontamination of equipment to avoid
cross-contamination. One of the oldest devices used to achieve emulsification through cavitation is
the liquid whistle. Process material is forced under pressure generated by a powerful pump through
an orifice from which it emerges and expands into a mixing chamber (Figure 9.5). With no moving
parts, other than a pump, the system is rugged and durable (Moser et al., 2001).
The systems that are particularly suitable for large-scale usage in the food industry are resonating
tube reactors. The liquid to be processed is passed through a pipe with ultrasonically vibrating
walls. In this way the sound energy generated from transducers bonded to the outside of the tube is
transferred directly into the flowing liquid. Generally, commercial tube reactors are constructed of
stainless steel. An alternative arrangement is through the coaxial insertion of a radially emitting bar
into the pipe containing the flowing liquid; which requires minimal change to existing pipework.
(a) (b)
Figure 9.4 (a) ultrasonic bath and (b) ultrasonic probe system.
Figure 9.5 Liquid whistle.
One such system consists of a hollow tube sealed at one end and driven at the other by a standard
piezo transducer. Another concept involves a cylindrical bar of titanium with opposing piezoelectric
transducers attached at each end. The design of both inserts is such that the ultrasonic energy is
emitted radially at half wavelength distances along their lengths.
9.3 appLICatIONS OF ULtraSONICS IN FOOD prOCESSING
The use of ultrasonics alone as well as its combination with other technologies was reviewed
by many researchers (Demirdoven and Baysal, 2009; Rastogi, 2011; Jimenez-Sanchez et al., 2017a,
2017b). Various types of ultrasonic systems are currently being used in food processing (Figure 9.6).
The application of ultrasonic waves in food processing includes extraction and separation, sterilization
and fresh-keeping, detection and analysis in food, freezing, thawing, crystallization, filtration,
drying, and osmotic dehydration (OD) (Ma et al., 2016). Some of these applications are discussed in
following sections.
Figure 9.6 ultrasonic systems used in food industry: (a) ultrasonic bath, (b) batch-type probe system, and
(c) continuous probe system. (From Zinoviadou et al., Food Res. Int., 77, 743–752, 2015.)
9.3.1 Ultrasonic for Microbial Inactivation
Ultrasonics alone or in combination with other hurdles is a promising alternative for the rapid
inactivation of microorganisms. The decimal reduction time values of resistance to ultrasonic
treatment (20 kHz, 200 kPa, 40°C) of various bacteria revealed that Salmonella was the least resis-
tant (0.80 min) and Bacillus subtilis was the most resistant (12 min). The sensitivity of ultrasonication
treatment was dependent on the type of organisms as D values for the microorganisms were differ-
ent, e.g., Aeromonas hydrophila (0.86 min), Escherichia coli (0.87 min), Pseudomonas aeruginosa
(0.92 min), Yersinia enterocolitica (1.2 min), Listeria monocytogenes (1.5 min), and Enterococcus
faecium (4 min) (Condon et al., 2004). The microbial inactivation during ultrasonic treatment is
mainly due to thinning of cell membranes, localized heating, and production of free radicals (Scheba
et al., 1991; Fellows, 2000; Butz and Tauscher, 2002). During the sonication process, regions of alter-
nating compression and expansion are created (Sala et al., 1995), which cause cavitation to occur, and
gas bubbles are formed in the medium. These bubbles collide violently during sonication cycles cre-
ating shock waves, which create regions of very high temperature and pressure. The pressure changes
resulting from these implosions are the main cause of bactericidal effect in ultrasonication, as the
hot zones created during sonication can kill only a few bacteria, that too are localized, and therefore
will not have an effect on the complete microbial population in food materials. The effectiveness of
microbial inactivation is dependent on the amplitude of the ultrasonic waves, exposure/contact time,
the volume of food being processed, the composition of the food, initial microflora of food, and the
treatment temperature (Piyasena et al., 2003).
Ultrasound (7.5 min, 40 kHz, 180 W) was applied to strawberry juice enriched with inulin and
oligofructose (Cassani et al., 2017) to enhance its storability, and it was observed that the nutritional,
microbiological, and sensory attributes of the product were better maintained in the treated juice.
Ultrasound was also demonstrated as a suitable non-thermal treatment for coconut water preserva-
tion due to its capacity to inactivate microbial population (Ribeiro et al., 2017). Ultrasonication
(24 kHz for 30 min) caused more than 4-log reduction of total plate count, yeast, and mould popu-
lation of mulberry juice (Engmann et al., 2015). Hosseinzadeh Samani et al. (2015) reported that
ultrasound power, ultrasonic exposure time, and temperature of the treatment were the most effec-
tive factors to reduce the microbial population (S. cerevisiae) in orange juice. Wang et al. (2015)
demonstrated that ultrasound (106.19 W/L) was found to be effective in reducing the spoilage micro-
organisms of cherry tomatoes. Ultrasound treatment (33 kHz, 60 W) for 40 min showed better
results during the cherry fruit storage as it showed significantly less microbial load than control
samples throughout storage (Muzaffar et al., 2016).
9.3.2 Ultrasonics in Combination with Other treatments for Microbial Inactivation
A combination of US treatment (24 kHz, 0.33 W/g, 60 min) along with heat treatment at 75°C
doubled the thermal inactivation rate of C. perfringens spores in beef slurry from a less than 1.5-log
reduction by ultrasound alone (Evelyn and Silva, 2015). A combination of ultrasound under pressure
and thermal treatment (mano-thermo-sonication) was shown to be more effective against bacterial
spore of B. mycoides, B. weihenstephanensis, and Psychrobacillus psychrodurans (isolated from
pasteurized edible crab meat) as compared to the individual treatments and showed a synergistic
effect on the inactivation of these spores (Condon-Abanto et al., 2016).
Ultrasonication in combination with thermal treatment (thermo-sonication) is a promising method
for the reduction of microbial spores. The pre-treatment with ultrasonication (20 kHz, 750 W) of
whole milk resulted in 35% reduction in D-value compared to thermal treatment at 100°C (Ansaria
et al., 2017). Thermo-sonication was reported to increase the microbial inactivation rate, extend
product shelf life, and reduce the impact on the nutritional content while maintaining the overall
quality of fruit and vegetable juices (Anaya-Esparza et al., 2017). The optimum thermosonication
conditions for prickly pear juice blend were 44°C–46°C for 19–20 min, which resulted in more than
3-log reduction of total plate count, enterobacteria, and yeast counts (Cruz-Cansino et al., 2016).
A higher microbial reduction (about 5-log reduction) of natural microbial flora in coconut water was
achieved for super critical carbon dioxide and ultrasound combination at 12 MPa and 40°C in about
15 min while about 30 min were needed for super critical carbon dioxide treatment alone. During
storage, super critical carbon dioxide-treated coconut water showed microbiological growth, while
samples subjected to combined treatment were safe for 4 weeks (Cappelletti et al., 2014).
Garud et al. (2017) observed that combination of ultrasound and thermal treatment (thermo-
sonication) was more effective as compared to the sonication alone. The time for 5-log reduction
of B. cereus and E. coil in various media (saline, broth, and sugarcane juice) in case of sonication
was found to reduce below 9 and 15 min, respectively, for the thermosonication as compared to the
11–31 min and 19–25 min, respectively, for ultrasonic treatment.
Combination of antimicrobial natural compound (pomegranate extract, 180 μg/mL) and sonication
(180 W, 40 kHz, 30 min) were shown to have a synergistic effect on kiwi fruit juice preservation
with higher reduction of yeasts and molds as compared to individual treatments (Tomadoni et al.,
2017). The combination of 2% cinnamon oil and ultrasound was shown to result in 0.85-log inhibition
of artificially inoculated L. monocytogenes in the mixtures of lettuce, parsley, and dill (Ozcan and
Zorba, 2016).
Ultrasound alone did not have much impact on A. acidoterrestris spores in commercial apple
juices, but the combination of ultrasound and ultraviolet (UV) treatment reduced spore count
significantly higher than that of the individual treatments (Tremarin et al., 2017). The combined
treatments of ultrasound with UV and UV with acid-electrolysed water showed a significantly higher
reduction in L. monocytogenes in raw salmon fillets compared to control (washed with sterile dis-
tilled water). The colour and odour of salmon were significantly affected by combined treatments,
but the texture and firmness of tissue did not change significantly (Miks-Krajnik et al., 2017).
The effects of ultrasound treatment (40 kHz, 500 W, 5 min) alone or in combination with
acetic acid, peracetic acid, or sodium dodecylbenzene sulfonate on storage stability of strawberry
(9 days at 8°C) showed that the ultrasound in combination with peracetic acid reduced the con-
taminating microbiota (moulds and yeasts, mesophilic aerobic, and lactic acid bacteria) without any
major physicochemical or sensory changes (Rosario et al., 2017). The combination of ultrasound
and organic acids (1% lactic acid) was found to be more effective as compared to the individual
treatments, as bacterial decontamination from green peppers and melons showed a higher removal
of pathogens by combined treatment. In case of green pepper, the combined treatment resulted in a
reduction of E. coli and S. enteritidis of the order of 2.8 and 2.9 log CFU/cm2, respectively, whereas
for melons, these values were 2.5 and 3.1 log CFU/cm2, respectively (Jose et al., 2014).
Luo et al. (2016) demonstrated that combination of ultrasound treatment (400 W/L, 3 min)
with moderate thermal treatment (40°C) significantly reduced B. cereus population (2.3 log
CFU/g) with minimal change in the colour of potato. A combination of slightly acid electrolyzed
water (pH 5.3–5.5; ORP 958–981 mV; ACC 28–30 mg/L) with simultaneous ultrasound treatment
at moderate temperature resulted in a higher reduction in B. cereus (approximately 3.0 log CFU/g).
Similarly, Luo and Oh (2015, 2016) demonstrated that the combined treatment (slightly acidic
electrolyzed water, ultrasound, and moderate thermal treatment of 60°C) of fresh-cut bell pepper
resulted in significant increase of lag time for L. monocytogenes and S. enterica, and therefore
prolonged the shelf-life of bell pepper (up to 8 days) at 4°C storage.
A combination of ultrasound (37 kHz, 380 W, 100 min) and NaOCl (200 ppm) treatments were
shown to have synergistic effects on C. sakazakii in fresh head lettuce (Park et al., 2015). Combined
ultrasound (15 min, 210 W) and near neutral electrolyzed (NEO) water (155 mg/l chlorine at pH 6.5)
treatment caused higher reduction of E. coli O157:H7 (4.4 log) and Salmonella Typhimurium DT
104 (4.3 log) inoculated on romaine lettuce leaves as compared to 2.8-log reductions for treatments
without ultrasound (Afari et al., 2015).
Inactivation of Botrytis cinerea, Penicillium expansum, Colletotrichum gloeosporioides, and

Alternaria alternata by ultrasonic fogging of dodecyl dimethyl ammonium chloride (500 ppm) was
achieved in 30 min (Daus et al., 2011), and fumigation at 20°C was considerably more effective than
fumigation at 5°C. The combined treatment of 200 ppm NaOCl and ultrasound (37 kHz, 380 W,
60 min) was found to be an optimum hurdle approach in the seaweed production, processing, and
distribution process to enhance seaweed safety (Park et al., 2014).
Ultrasonication has very small effect on cell density up to a power level of 40 W, but at and
above 60 W, a dramatic decrease in cell density was observed probably due to the cell lysis. Efficacy
of low-power ultrasonication could not be enhanced even by combining with antimicrobial peptide
(melittin) at low concentration to inactivate L. monocytogenes (Wu and Narsimhan, 2017). A few
selected studies on microbial inactivation by ultrasonication alone or in combination with other
treatments are presented in Table 9.1.
table 9.1 Effect of Ultrasonication alone or in Combination with Other treatments on the Growth of
Microorganism in Various Juices
S. No. product Ultrasonic Conditions Effect on Microbes reference (s)
1 orange juice 24 kHz, 78°C, 60 min 4.4-log reduction of evelyn and silva
(juice pretreated at Alicyclobacillus acidoterrestris (2016)
600 m Pa for 15 min) spores
200 W, 24 kHz, 30 min 2.8-log reduction of sanchez-rubo et al.
Saccharomyces cerevisiae (2016)
24 kHz, 50°C, 5 min 3.37-log reduction of Escherichia Munoz et al. (2011)
coli
20 kHz Combination with uV caused Char et al. (2010)
inactivation of Escherichia coli
and Saccharomyces cerevisiae
30 kHz, 55°C, 30 min 5.5-log reduction of Walkling ribeiro
Staphylococcus aureus et al. (2009)
vegetative cells
600 W, 20 kHz Combination with mild heat Ferrante et al.
treatment (45°C), and natural (2007)
antimicrobials (vanillin 1,000 ppm
and citral 100 ppm) was effective
for the control of Listeria
monocytogenes
2 Apple juice 24 kHz, 75°C, 30 min 1.6-log reduction of Neosartorya evelyn et al. (2016)
fischeri ascospores
20 kHz, 44°C, 30 min 6.4- and 5.8-log reduction of Ferrario et al. (2015)
Saccharomyces cerevisiae, and
3.0- and 2.0-log reduction of
Alicyclobacillus acidoterrestris
spores in commercial juice and
natural juice, respectively
20 kHz, 56°C, 0.28 min 1-log reduction of Chromobacter Arroyo et al. (2012)
sakazakii
24 kHz, 50°C, 2.9 min 4.9-log reduction of Escherichia Munoz et al. (2012)
coli
24 kHz, 60°C, 30 min 7-log reduction of Saccharomyces Marx et al. (2011)
cerevisiae
45 kHz, 50°C, 30 min 4.56-log reduction of Wang et al. (2010)
spores
20 kHz inactivation of Escherichia coli Patil et al. (2009)
300 W, 60 min 90% inactivation of yuan et al. (2009)
(Continued)
table 9.1 (Continued) Effect of Ultrasonication alone or in Combination with Other treatments on the
Growth of Microorganism in Various Juices
S. No. product Ultrasonic Conditions Effect on Microbes reference (s)

Apple cider 48 kHz 5-log reduction was achieved for rodgers and ryser
L. monocytogenes and (2004)
Escherichia coli o157:H7 (with
proper sanitation protocol before
juice extraction)
3 Pomegranate 200 W, 24 kHz, 50°C, 2.52-log reduction of sanchez-rubo et al.
juice 30 min Saccharomyces cerevisiae (2016)
4 tomato juice 20 kHz, 10 min yeast Adekunte et al.
(2010c)
20–24 kHz, total lactic acid bacteria Adekunte et al.
26 s–20.4 min (2010a, 2010b)
20–24 kHz, Fusarium oxysporum, Salmonella bevilacqua et al.
26 s–20.4 min, spp., Shigella spp. (2013)
benzoate (100 ppm)
and citrus extract
(1800 ppm)
5 Mango juice 25 kHz, 60°C, 7 min 5-log reduction of Escherichia coli Kiang et al. (2013)
7-log reduction of Salmonella
enteritidis
6 Pineapple 24 kHz, 60°C, 6 min 5-log reduction of Saccharomyces bermudez-Aguirre
juice cerevisiae and barbosa
Canovas (2012)
7 sugarcane 20 kHz, 50°C, 10 min 5-log reduction of Escherichia coli Garud et al. (2017)
juice 2.5-log reduction of Bacillus
cereus
5-log reduction of natural
microflora predicted in 29 min
9.3.3 Ultrasonic-assisted Enzyme Inactivation
Ultrasonication has a potential to overcome harmful effects of thermal processing with retention
of phytonutrients and, therefore, offers a better alternative in various food processing applications.
The cavitation and micro-streaming created by ultrasonic treatment generate thermal, mechanical,
and chemical effects to inactivate enzymes. The cycles of pressurization and depressurization have
mechanical effect besides causing localized heating. Hydrogen bonds can be broken and the van
der Waals interactions in polypeptide chains may be weakened by the formation and collapse of
bubbles. Ultrasonication may alter the secondary and tertiary structure of enzymes resulting in the
loss of activity (Islam et al., 2014a). The inactivation mechanisms of the enzyme by ultrasound in
food processing were recently reviewed by Cheng et al. (2016).
The complete inactivation of bael peroxidase was achieved by thermal processing (85°C, 11 min)
and the retention of marmelosin was 16.71%, while ultrasound treatment (64.07 Wcm−2, 4 min)
resulted in complete enzyme inactivation with higher retention of marmelosin (49.8%) (Mugdha and
Ananthanarayan, 2017). Ribeiro et al. (2017) showed that ultrasound had an additive effect to the heat
treatment on the inactivation of the polyphenol oxidase (PPO) and peroxidase (POD) of coconut water.
Ultrasonication (24 kHz, 30 min) also reduced PPO and POD activities of mulberry juice (Engmann
et al., 2015). Variable inactivation of POD and PPO in apple cubes and apple juice was achieved by ultra-
sonic processing (55–3300 W/L) at 23°C (ambient) to 60°C for 5–20 min (Silva et al., 2015). PPO activity
in the pineapple juice was reduced to 20% by ultrasonic treatment (376 W/cm2 and 10 min) as compared
to the control, and it enhanced the juice colour and the product was stable for 42 days of storage (Costa
et al., 2013). Similar ultrasonic treatment on cantaloupe melon juice resulted in significant reduction of
POD and PPO, and the total inactivation of ascorbate peroxidase (APx) (Fonteles et al., 2012).
Ultrasonication (200 W, 15 min) of egg yolk prior to incubating it with cholesterol oxidase
(0.6 U/g egg yolk) for 10 h at 37°C resulted in 91.68% reduction of cholesterol level in egg yolk
without affecting the quality attributes of the yolk indicating that the ultrasonication can be used
to enhance degradation of cholesterol in egg yolk (Sun et al., 2011). Pectin methylesterase (PME)
activity of prickly pear juice was reduced by 54% as compared to control by ultrasound treatment
(Cruz-Cansino et al., 2016). Recent reports on enzyme inactivation by ultrasonication alone or in
combination with other treatments are compiled in Table 9.2.
table 9.2 Effect of Ultrasonication alone or in Combination with Other treatments on Enzyme activity
in Various Foods
Ultrasonic
SN product Conditions Effect on Enzymes reference (s)
1 orange juice 24 kHz, 63°C, 91% inactivation of PMe Koshani et al. (2014)
9.8 min
30 kHz, 55°C, 20 min 96% inactivation of PMe Walkling-ribeiro
et al. (2009)
20 kHz, with acoustic 62% inactivation of PMe tiwari et al. (2009)
density of 1.05 W/mL,
10 min
2 tomato juice 20 kHz, 75°C, 4 min Complete inactivation of PMe and terefe et al. (2009)
72% inactivation of
polygalacturonase
24 kHz, 65°C, 8 min 98.9% inactivation of PMe Wu et al. (2008)
24 kHz, 70°C, 90% inhibition of PMe Wu et al. (2008)
4.3 min
20 kHz, 100 W, D value for inactivation of PMe raviyan et al. (2005)
20 min reduced from 24 min for
ultrasonication to 0.3 min for
thermosonication at 72°C
tomato paste 20 kHz, 70°C, 1 min Complete inactivation of PMe Vercet et al. (2002)
62% inactivation of polygalacturonase
3 Pineapple 19–20 kHz, 376 W/cm2 20% reduction in polyphenol oxidase Costa et al. (2013)
juice and 10 min activity
4 Cantaloupe 19–20 kHz, 376 W/cm2 significant reduction of PoD and Fonteles et al. (2012)
melon juice and 10 min polyphenol oxidase and the total
inactivation of ascorbate PoD
5 Carrot 20 kHz, 60°C, 10 min More than 90% inactivation of PMe, Jabbar et al. (2015b)
polyphenol oxidase, and PoD
6 Grape 28 kHz, 60°C, 60 min 91% inactivation of PMe Aadil et al. (2015)
90% inactivation of polyphenol
oxidase
89% inactivation of PoD
7 Apple 20 kHz, 60°C, 10 min 92.9% inactivation of PMe Abid et al. (2014)
93.85% inactivation of polyphenol
oxidase
91% inactivation of PoD
8 Fresh cut 40 kHz synergistic effect on inactivation of Jang and Moon
apple polyphenol oxidase and PoD when (2011)
combined with ascorbic acid (1%)
9 Guava 20 kHz Higher inactivation of PoD when Ganjloo et al. (2008)
combined with heat (80°C–95°C)
10 Watercress 20 kHz, 50% power Higher inactivation of PoD than Cruz et al. (2006)
juice blanching at 85°C
9.3.4 Ultrasonic-assisted Extraction of Biomolecules
Use of ultrasound as the green approach in the processes of preservation, transformation, and extrac-
tion of valuable materials has been described recently (Chemat et al., 2017). The ultrasonication has
been used to increase the extraction yield and improve the quality of extracts. It is a simple, quick, and
inexpensive as well as environment friendly technique. Application of ultrasound-assisted extraction
(UAE) showed significantly higher recovery of phenolic compounds compared to solid-liquid extrac-
tion process alone from peels of potato with lower ultrasonic frequency (33 kHz) being more effective in
recovering polyphenols (Kumari et al., 2017). UAE of anthocyanins (23.54 mg Cyn-3-Glu/100 g) from
the pericarp of mangosteen seed was achieved in 5 min against 4 h of shaking water bath extraction
(Hiranrangsee et al., 2016). UAE of alginates from brown seaweeds (Sargassum binderi and Turbinaria
ornata) and carrageenans from red seaweeds (Kappaphycus alvarezii and Euchema denticulatum) was
achieved in a shorter time (15–30 min) as compared to 2 h of conventional extraction, and UAE was
found to retain chemical properties of extracted compounds (Youssouf et al., 2017).
At the optimal extraction conditions for UAE (0.40 W/mL, temperature 60°C, solid-liquid ratio
1:50 g/mL), the yield of pectin from grapefruit peel was higher (26.74%) and extraction time was
shorter (51.79 min) as compared to conventional heating extraction (80°C, 60 min) (Xu et al., 2014c).
The application of UAE (144 W, 70°C, 50 min) resulted in higher yield of the pigment from red rice
(16.66%) at the optimal conditions (Lin et al., 2011). Fernandez-Ronco et al. (2013) observed that
although extraction yields of Capsicum annuum oleoresin were not affected by the extraction tech-
nique, the UAE reduced the extraction time significantly. Ultrasound treatment was applied to olive
paste before mechanical extraction, which resulted in a virgin olive oil that had a more harmonic
taste and an appealing and desirable colour than the traditional one (Clodoveo et al., 2013).
UAE has several advantages such as energy efficiency and time-saving. The recent reports of
UAE include extraction of carotenoids (Jin and Ma, 2017), oleoresins (Melgar-Lalanne et al., 2017),
red and yellow pigments (Zhang et al., 2014), polysaccharides (Wu et al., 2014), phenolics (Cavuldak
et al., 2016), and bioactive compounds and antioxidants (Jabbar et al., 2015a). The optimized UAE
conditions for some of the bioactive compounds are presented in Table 9.3.
9.3.5 Ultrasonic-assisted Drying
Ultrasound has been used to accelerate solid gain and shorten drying time. A faster drying of
vegetables was achieved with high-intensity ultrasound in combination with hot air systems even at
lower temperatures as compared to hot air drying alone (Gallego-Juarez et al., 2007), and the product
quality was found to be superior (Garcia-Perez et al., 2007). Designs of dryers involving ultrasound
to enhance the drying process and various possible mechanisms of accelerated drying process were
recently reviewed by Musielak et al. (2016).
The use of ultrasound in combination with OD resulted in higher rate of water loss and sol-
ute gain at a lower solution temperature while preserving the natural flavour, color, and heat-
sensitive nutritive components. The rehydration of ultrasonically treated samples showed that
the percentage of rehydration was higher as compared to that of untreated samples for many
fruits. Sour cherries were osmo-dried using a 60% sucrose solution for 120 min (40°C) during
which ultrasound (25 kHz, 0.4 W/cm 2) was applied, and it was further dried in the convective
dryer, which resulted in faster drying rate as compared to control (Siucinska et al., 2016). Apple
samples subjected to ultrasonic treatment (21 and 35 kHz for 30 min) prior to drying in a con-
vective oven at 70°C showed a reduction in drying time by 13%–17% (Fijalkowska et al., 2016).
Ultrasound (35 kHz) prior to drying of apple cubes by convection method (70°C, air velocity
1.5 m/s) resulted in the reduction of the drying time by 31%, and ultrasound-treated apples exhib-
ited higher shrinkage, lower density, and higher porosity than untreated samples (Nowacka et al.,
2012). The air drying of ultrasonically pre-treated banana and melon fruits resulted in an increase
table 9.3 Optimized Ultrasound-assisted Conditions for Extraction of Various Bioactives
156
Ultrasonic Solid-liquid Bioactive Extracted

S. No. plant Ultrasonic power time Extraction Condition ratio (g/mL) and Yield reference (s)
1 Lingonberry 450 W 25 min 50% ethanol as solvent 1:15 total flavonoids, 29.87 mg/g Li et al. (2017)
2 Althea 86.44 W 36.86 min temperature of 79.93°C 1: 39.99 Crude polysaccharides, 12% Pakrokh Ghavi
officinalis (2015)
roots
3 red pepper 300 W 30 min 95% ethanol as solvent 1:12 Capsanthin, 19.22% Meng and
temperature of 30°C Ming-hua
(2017)
4 Opuntia ficus 40 kHz, 330 W 70 min temperature of 70°C, pH 1.5 1: 30 Pectin,18.14% bayar et al.
indica (2017)
5 Apricot kernels 40 kHz 43.95 min temperature of 51.72°C 1: 19.8 oil, 46.01% Gayas et al.
(2017)
6 Chia seed 40 kHz, 176 W 40 min temperature of 50°C ethyl 1:12 oil, 27.24% Mello et al.
acetate as solvent (2017)
7 Chlorella 83.7% amplitude 7.03 min 62.03% ethanol as solvent 1:100 β-carotene, 55 μg/g singh et al.
(2017)
8 Ocimum 0.26 W/cm3 11.71 min 55.34% ethanol as solvent 1:40 Flavonoids, 6.69 mg of upadhyay et al.
tenuiflorum quercetin equivalents/g; and (2015)
leaves phenolics, 9.41 mg of gallic
acid equivalents/g
9 Artichoke 20 kHz, 240 W 60 min 50% ethanol as solvent 1:10 Chlorogenic acid, 16.47 mg/g rabelo et al.
dry matter (2016)
10 sorghum shell 0.32-Wcm−2 10 min temperature of 50°C, 1:15 Phenolics, 35.66 mg re/g Hou et al. (2016)
ultrasonic intensity 80% ethanol as solvent
bath extraction was done twice
11 Trapa 40 kHz, 200 W 41 min temperature of 58°C 1:31.5 Crude polysaccharides, raza et al.
quadrispinosa 2.78% (2017)
stems
12 Glycyrrhiza 270 W 6h — 1: 20 essential oils, 0.209% Dou et al. (2016)
13 Walnut leaves 35 kHz, 480 W 51.28 min 61% ethanol as solvent 1: 4.96 total phenolics, 10.125 g nour et al.
gallic acid equivalents (2016)
per kg
14 Walnut leaves 35 kHz, 480 W 49.37 min 67.83% ethanol as solvent 1: 4.96 total flavonoids, 2.925 g nour et al.
quercetin equivalents per kg (2016)
(Continued)
table 9.3 (Continued) Optimized Ultrasound-assisted Conditions for Extraction of Various Bioactives
Ultrasonic Solid-liquid Bioactive Extracted
S. No. plant Ultrasonic power time Extraction Condition ratio (g/mL) and Yield reference (s)
15 Pueraria lobata 40 kHz, 3.0 W/cm 2 39.79 min Acid ethanol as solvent (HCl 1:10 Puerarin, 43.04 mg/g Zou et al. (2016)
root 2M/L) at temperature of
57.82°C
16 nutmeg 40% of maximal 10 min 100% ethanol as solvent at 1:4 oleoresin, 9.63% Morsy (2016)
power room temperature
17 Quince seed 24 KHz probe and 30 min temperature of 47°C 1: 32.5 Mucilage, 42.7% Farahmand
400 W power Water as solvent et al. (2016)
18 Opuntia 20 kHz, 130 W 10 min 40% methanol as solvent 1: 20 total phenols, 2663.4 mg espinosa-
oligacantha GAe/100 g DW Munoz et al.
(2017)
19 Opuntia 20 kHz, 130 W 20 min 40% methanol as solvent 1: 30 Flavonoids, 17.84 mg espinosa-
oligacantha Qe/100 g DW Munoz et al.
(2017)
APPLiCAtion oF uLtrAsoniC in FooD ProCessinG
20 Artemisia 150 W 14.5 min temperature of 60°C 1:25 Polysaccharides, 8.86% Wang et al.
selengensis (2016)
leaves
21 rapeseed meal 252.9 W 48.5 min temperature of 49.6°C 1:41.4 Carotenoids, 0.1577 mg/g Fengwei et al.
(2015)
22 red cabbage 30 kHz, 100 W 30 min temperature of 15°C 1:50 Anthocyanins, 20.9 mg/Kg ravanfa et al.
Water as solvent (2015)
23 red cabbage 37 kHz 75 min temperature of 40°C 1:3 11.92% higher anthocyanin Demirdoven
42.39% ethanol as solvent than conventional process et al. (2015)
24 Cassia 50 W 5 min solvent concentration 60%, 1: 25 total phenolics, 59.68 mg sharmila et al.
auriculata pH 6.2 GAe/g (2016)
leaves
25 Dried jujube ultrasonic frequency 50 min six times ex 1: 12 Azocyclotin, 0.15 mg/kg tian et al.
of 45 kHz, an extraction at 45°C (2015)
ultrasonic power of
70 W
157
in effective water diffusivity during air drying leading to a reduction in the drying time by 25%
(Fernandes et al., 2009). Azoubel et al. (2010) indicated that the application of ultrasound during
drying of banana could increase the moisture diffusivities, which reduced process time leading to
the saving of energy. The ultrasound treatment also reduced the drying time without affecting
the product quality in terms of bulk density, colour, ascorbic acid content, and rehydration char-
acteristics for vegetables (Schossler et al., 2012).
The application of ultrasound in dehydration and OD was reviewed by Mason et al. (2005). The
effects of ultrasound-assisted OD combined with hot air drying of carrot slices showed that ultra-
sonic energy density and solution concentration have significant effects on weight reduction ratio.
OD pre-treatment without ultrasound assistance before hot air drying increased the total processing
time and reduced the effective moisture diffusivity. When ultrasound was applied, the increase of
ultrasonic energy density reduced both dehydration time and total processing time. Ultrasound
prior to hot air drying had positive effects on improving the carotenoid content of the product and
reducing process energy cost (Liu et al., 2014).
9.3.6 Ultrasonic-assisted Freezing
Ultrasound has been used to assist the freezing process of both solid and liquid samples (Xu et al.,
2015). Ultrasonic-assisted freezing (UAF) for food is a relatively new and promising application to
enhance the freezing process and preserve food quality (Li and Sun, 2002; Sun and Li, 2003; Zheng
and Sun, 2006; Delgado et al., 2009; Delgado and Sun, 2011; Islam et al., 2014b; Xu et al., 2014a; Cheng
et al., 2017). This enhancement was achieved by initiating ice nucleation, increasing heat and mass
transfer rates, and controlling the size and shape of ice crystals (Acton and Morris, 1992; Chow et al.,
2003, 2004, 2005). Ruecroft et al. (2005) and Saclier et al. (2010) also reported that power ultrasound
could initiate nucleation, and the nucleation occurs immediately once the cavitation bubbles collapse.
Furthermore, the pressure gradient generated by cavitation bubbles is also regarded as the driving force
for ice nucleation (Dodds et al., 2007; Grossier et al., 2007). In addition, micro-streaming can also be
considered as a driving mechanism for the ice nucleation during UAF process (Zhang et al., 2003).
Ultrasound (20 kHz) treatment of radish samples at −0.5°C for 7 s duration with the intensity
of 0.26 Wcm−2 was able to induce nucleation, and it significantly shortened the time of dehydration
and subsequent freezing of radish. The ultrasound-assisted freezing (UAF) process with 25 kHz
frequency at 288 or 360 W power levels shortened the total time for dough freezing by 11%, and
enhanced the nucleation, as the formation of a larger number of tiny ice crystals inside the frozen
dough was observed as compared to control dough (Hu et al., 2013).
Compared to OD, ultrasound-assisted osmotic dehydration (UOD) products exhibited less freez-
able water and better firmness and microstructure. After freezing/thawing, frozen products of UOD
also displayed lesser drip and ascorbic acid losses and maintained better firmness than that of OD
samples (Xu et al., 2014b). Similarly, nucleation in the centre of potato cubes was induced by ultra-
sound (35 kHz) treatment at −2.0°C (Comandini et al., 2013), and the enhanced nucleation resulted
in reduced freezing time. Sun and Li (2003) examined the microstructure of potato tissues frozen
by ultrasound-assisted immersion freezing at 25 kHz and 15.85 W. The results showed that not only
the freezing rate was increased but also the occurrence of cell destruction was avoided by UAF.
In strawberry also, the power ultrasound increased the nucleation and shortened the freezing time.
The freeze-dried sample frozen by UAF had finer cellular structure compared to those frozen in
other freezing conditions (Cheng et al., 2014). Delgado et al. (2009) indicated that ultrasound-assisted
immersion freezing of apple samples enhanced the freezing rate up to 8% as compared to control,
which may be due to the induction of primary nucleation by ultrasound. Microstructure and the
firmness of broccoli tissue frozen by UAF (30 kHz) were markedly better than those of broccoli
tissue frozen by traditional immersion freezing method, and the drip loss of samples frozen by
UAF was significantly reduced (Xin et al., 2014b). The UOD was found to preserve the firmness
and minimize the drip loss and loss of ascorbic acid content in osmodehydrofrozen broccoli during
frozen storage at −25°C for 6 months (Xin et al., 2014a).
The resistance of heat and mass transfer was reduced by ultrasonication at the ice/liquid
interface leading to the enhancement in the freezing rate (Li and Sun, 2002; Sun and Zheng, 2006).
Cavitation clouds at the surface of the sphere sample have the main effect of enhancing heat transfer
(Kiani et al., 2012), and high rate of heat and mass transfer can enhance ice crystal growth. Many
factors affect the effectiveness of UAF, such as the intensity and frequency of ultrasound, the posi-
tion of the samples, cooling medium temperature, and flow rate. The properties of the cooling
medium could also significantly affect heat transfer and ice crystal formation. Zheng and Sun
(2005, 2006) reviewed the application of ultrasonics in the freezing process, and the mechanism of
ultrasonic-assisted refrigeration was recently described by Pang et al. (2017).
9.3.7 Ultrasonic-assisted Improvement in animal product Quality
Ultrasonication has been suggested as a new promising technique to improve the quality of animal
products. Saleem and Ahmad (2016) reported that the ultrasonication at low frequency (20 kHz) can
improve functional properties of meat and meat products. Lei et al. (2016) also reviewed the effect of
ultrasound on meat quality and its prospect in the meat industry. Ultrasound is a useful tool for the
meat industry as it helps in tenderization, acceleration of maturation and mass transfer, reduction in
cooking energy, and increase in shelf life of meat without affecting other quality parameters. It also
improves functional properties of emulsified products, eases cleaning, and is used for sterilization of
equipment surfaces (Alarcon-Rojo et al., 2015).
The gelation characteristics of myofibrillar protein are largely responsible for the textural and
functional properties of fish products and are especially important for the quality of surimi. The gel
made up of Ctenopharyngodon idellus myofibrillar protein treated with ultrasonics at 600 W gave
the best gel properties, such as the highest gel strength, hardness, gumminess, chewiness, WHC,
and whiteness. The reduced particle size, narrowed particle distribution, decreased free SH con-
tent, and intrinsic fluorescence, as well as improved surface hydrophobicity caused by US treatment,
indicated the unfolding and disaggregation of myofibrillar protein, which also improved the gel
properties (Wen et al., 2017). The ultrasound-assisted hydrolysis (70 W) of tilapia (Oreochromis
niloticus) muscle protein caused a reduction in the degree of hydrolysis ranging from 23% to 35%
relative to that of the conventional process and increased antioxidant activity in comparison to
control (Sureeporn et al., 2014).
Ultrasonication of fresh chicken in marinating liquid for 40 to 60 min caused significant
changes in the rate of curing absorptivity, shear force, cooking loss, and NaCl content probably due
to high molecular weight proteins decomposition by the ultrasound treatment (Feng et al., 2014).
Ultrasonication (10, 25, or 40 min; 4.2, 11, or 19 Wcm−2) was also found to decrease cohesiveness
and gumminess of meat samples, and NaCl content increased significantly in all sonicated samples
indicating that ultrasonic curing can assist in brine infusion, reducing processing times with minimal
impact on product quality (McDonnell et al., 2014). Various meat tenderization methods, including
physical methods (electrical stimulation, high-pressure processing, and ultrasound), biochemical
methods (salts, organic acids, and enzyme), processing techniques, and combined methods were
reviewed by Wu et al. (2012). The use of ultrasound to improve meat tenderness was also described
as promising technology by Alves et al. (2013). Ultrasound treatment of pork (40 kHz, 150 W in
water, and 2% w/v brine at 4°C) resulted in the rapid thawing of pork with thawing rates similar
to those thawed at 17°C. Ultrasound-treated pork in brine had an advantage of less cooking losses;
however, a slight discoloration of the pork was noticed (Hong et al., 2014).
Ultrasonic treatment was found to result in increased protein concentration and a higher degree of
hydrolysis of eggshell membrane proteins hydrolysate as compared to the control samples (untreated
enzymatic hydrolysates). The functional properties of hydrolysate such as solubility, emulsification,
foaming properties, and water holding capacity were found to be improved by ultrasound treatment
(Jain and Anal, 2016). The power ultrasound (20 kHz, 20 min) improved the functionality of egg
white proteins (10% w/w aqueous solutions at pH 6.5-7.1) by enhancing its antioxidant and antimi-
crobial activities (Stefanovic et al., 2014a). Ultrasound (20 kHz, 4.27 W, and 20% of amplitude for
20 min) treatment of egg white proteins showed a decrease in the consistency index but surface
hydrophobicity was greatly increased; however, gelation performance and sulfhydryl content were
unchanged after ultrasound treatment (Arzeni et al., 2012). Stefanovic et al. (2014b) reported that
the ultrasound treatment (35 and 40 kHz) under controlled conditions (temperature, 25°C and 55°C;
time of pre-treatment, 15–60 min; pH, 7.00–10.00) can improve the hydrolysis of egg white proteins
solutions and improve their functional properties.
The effects of ultrasound on the structures, gelation properties, particle sizes, viscosity,
solubility, turbidity, emulsifying properties, and forming properties of food proteins as well as pro-
tein reaction, extraction, and sensory quality were reviewed (Hu et al., 2015). Ultrasound possesses
the capability to improve the functional properties of proteins by improving the surface-active
properties, which decreases protein aggregate size and increases molecular mobility. Enhancement
of enzymatic hydrolysis by ultrasound application can modify protein conformation by affecting
hydrogen bonds and hydrophobic interactions, disrupting the quaternary and/or tertiary structure of
proteins due to the effects of cavitation (Ozuna et al., 2015). Ultrasound (20 kHz, 4.27 W, and 20%
of amplitude, 20 min) promoted a decrease in the consistency index and changed the functionality
of whey protein concentrate as surface hydrophobicity was greatly increased after treatment (Arzeni
et al., 2012). Ultrasonication (20 kHz) increased the clarity of whey protein concentrate solutions
largely due to the reduction in the size of the suspended insoluble aggregates (Zisu et al., 2011). The
gel strength of whey protein concentrate solution when heated at 80°C for 20 min also increased
with sonication, while gelation time and gel syneresis were reduced. The microstructure of the whey
protein gels indicated a compact network of densely packed whey protein aggregates arising from
ultrasound treatment.
The solution of sodium caseinate, whey protein isolate, and milk protein isolate were sonicated
for 2 min with a power intensity of ~34 Wcm−2 to reduce the size and hydrodynamic volume of
the proteins without affecting their molecular weight. Emulsions produced with ultrasound-treated
sodium caseinate and whey protein isolate had the same droplet sizes as untreated proteins at all
concentrations, despite the reduction in protein size of the sonicated proteins (O’Sullivan et al., 2014).
The ultrasound pre-treatment (800 W, up to 20 min) of goat milk before rennet-induced coagulation
showed an increase in the degree of whey protein denaturation, while soluble calcium and phosphorus
were also increased. The gel firmness, coagulum strength, cohesiveness, water holding capacity, and
cross-linking of gels demonstrated marked increase, and the improvement in goat milk coagulation
properties was observed with increasing duration of ultrasound treatment (Zhao et al., 2014).
9.4 MISCELLaNEOUS appLICatIONS OF ULtraSONICS IN FOOD INDUStrY
The low-frequency and high-intensity ultrasound, which is also termed as power ultrasound, can
be used in different areas of food processing, such as blanching, emulsification/homogenization,
sterilization/pasteurization, extraction, drying, and freezing (Li et al., 2004; Hengl et al., 2014).
Other applications of ultrasound processes in food manufacturing include peeling, disintegration of
cells, activation (acceleration) of an enzyme reaction in liquid foods, acceleration of a microbial fer-
mentation, mixing, homogenization, emulsification of oil/fat in a liquid stream, spraying, degassing,
crystallization of fats and sugars, foam breaking, effluent treatment, humidifying, and fogging.
Utilization of low-power ultrasound in high frequencies as a non-destructive measuring tech-
nique is referred as analytical ultrasound. Propagation of low-power ultrasound in dairy products
provides information about physical, mechanical, and chemical properties of the product and the
changes generated through manufacturing processes. Wave velocity, attenuation, time of flight,
and impedance are the ultrasonic factors which are affected by the changes of compositional and
rheological properties of the medium. Analytical ultrasound has been applied in dairy products
for detecting microbial contamination, tracking coagulation process, detecting foreign bodies, and
textural and compositional characterization (Mohammadi et al., 2017).
Ultrasound has been shown to be a rapid, non-destructive technique for the detection of foreign
bodies (glass, metal, plastic), internal structural defects such as holes or cracks, and suspended par-
ticles in foods as compared to currently used techniques (optical inspection, metal detection, and
X-rays) in the food industry. Ultrasound-based techniques were reported to detect and identify for-
eign bodies in food products such as cheese and poultry (Benedito et al., 2001; Cho and Irudayaraj,
2003; Correia et al., 2008); bottled beverages (Zhao et al., 2003); and yogurt, fruit juices, and
tomato ketchup (Knorr et al., 2004).
Ultrasound can be used to determine the thickness of materials such as the thickness of choco-
late layers on confectionery, fat, or lean tissue in meat, liquids in cans and egg shells, particle
sizes in emulsions or suspensions, etc. Ultrasound can be used to measure sugar concentration
of aqueous solution, salt concentration of brine, triacylglycerols in oils, droplet concentration of
emulsions, alcohol content of beverages, composition of milk, and physical properties of batters
(density, viscosity, and rheology) and cakes (volume, symmetry, volume index, height, and density)
(McClement, 1995; Gomez et al., 2008).
The optimal ultrasound blanching conditions of green beans were found to be 90°C for 58.27 s
with duty cycle of 0.79 s as it resulted in the least residual POD activity (9.64%) and highest vitamin
C retention (91.08%), and it was found to be more efficient compared to the conventional blanching
method as it caused less damage to the product (Yolmeh and Najafzadeh, 2014). The ultrasound
technology can be used to improve the physical properties of peach juices, as it increases the
stability to pulp sedimentation, serum cloudiness, and juice consistency, with insignificant colour
changes during the storage (Rojas et al., 2016).
Ultrasound-assisted cutting has the potential to replace traditional cutting methods due to advan-
tages such as high accuracy, low product loss, less deformation, reduced friction, less down time, and
ability to handle sticky or brittle foods. The quality of cheeses (Cheddar, Mozzarella, and Swiss) cut
with an ultrasonic knife at three amplitudes (30%, 40%, and 50%) exhibited a relatively shiny and
smooth surface compared with the relatively dull and rough surfaces of the samples cut without ultra-
sound, and a better quality was observed when the ultrasound amplitude was increased from 30%
to 50%. The cheeses cut with ultrasound exhibited lower peroxide values compared to the control,
indicating less lipid degradation (Yildiz et al., 2016). The ultrasonic cutting can be applied to cut a
thin slice of food, as it has advantages of the excellent cut surface and reduced crumbling, squeezing,
debris, and smearing, which is not possible in conventional cutting methods (Liu et al., 2015).
The ultrasound (2.5 kW for 9 min)-assisted mixing was used to improve batter characteristics as
it lowered batter density and flow behaviour index, which resulted in higher overrun and viscosity
compared to the control, and produced better quality cake (lower hardness but higher springiness,
cohesiveness, and resilience) (Tan et al., 2011). Ultrasound can also be used to preserve the quality
of functional ingredients, such as fructo-oligosaccharides (FOS), which are known to be hydrolysed
and lose their functional properties when subjected to thermal processing. Filho et al. (2016) reported
that FOS solution (70 g L−1) subjected to ultrasound at 600–1200 W L−1 for 5 min did not show any
significant change on FOS concentration (<2.0%) or on the degree of polymerisation of the FOS (<3.3%).
Similarly, the hydrolysis of dextran under ultrasound, microwave, and irradiation shock combination
treatment was found to be significantly higher than under individual treatments (Bashari et al., 2016).
Ultrasonic processing has been used for generation of Maillard reaction products (MRP), and
the ultrasonic MRP were found to have higher depletion rates of reactants, higher generation rates of
intermediate, and the generation of MRP at relatively low processing temperatures (55°C and 60°C)
compared with those induced thermally (Yu et al., 2016). Ultrasonic pre-treated (200 W) soybean
protein isolate MRPs had a higher degree of glycation, lower browning, and less compact tertiary
conformation than those of non-ultrasonic pre-treated controls (Zhao et al., 2016). Power ultrasound
also improved the oxidative stability of the potato chips by 21% at the end of 8-week storage at 35°C
by removing some of the lipids from the surface (Wambura and Yang, 2011).
The ultrasound applications in dairy processing include emulsification, crystallisation, inacti-
vation of microbes, functionality modifications, and fat separation (Russell et al., 1998; Jayani and
Leong, 2015). The ultrasound-assisted mixing of liquid cheese aroma in different carbohydrate matri-
ces resulted in a lower microcapsule size and higher aroma retention as compared to mechanical mix-
ing prior to encapsulation by spray drying (Mongenot et al., 2000). The application of ultrasound also
prevented the fouling of milk in a double pipe heat exchanger due to the reduction of temperature at
the solid-liquid interface as well as the induced movement of the deposits (Lin and Chen, 2007). Use
of ultrasound reduced scaling and increased cleaning speed in the case of evaporation of sugar juice in
a tubular evaporator (Lu et al., 2005). The ultrasound-assisted soaking and gelatinization of rice (75°C
with sonication) resulted in a reduction in rice parboiling time (70%) as the moisture content of 48%
was achieved after 3 hours in combined treatment as compared to 10 hours in conventional soaking
(Wambura et al., 2008).
Application of ultrasound to membrane processes increased the flux by disturbing the concentration
polarization at the membrane surface. The liquid jet and cavitation mechanisms led to particle release
from the fouled membrane (Kyllonen et al., 2005). Ultrasound treatment increased the permeate flux
due to cavitation and acoustic streaming in case of membrane distillation (Zhu and Liu, 2000; Narayan
et al., 2002) and forward osmosis (Chanukya and Rastogi, 2017). Ultrasound effectively improved the
performance of dead-end ultrafiltration due to the removal of part of the boundary layer from the mem-
brane surface affecting the concentration polarization phenomenon (Simon et al., 2000). Application of
ultrasound increased the ultrafiltration flux as well as lysozyme rejection in cross-flow ultrafiltration of
binary bovine serum albumin and lysozyme (Tenga et al., 2006). Ultrasonic waves may also help to
displace the cake on the membrane structure by cavitation, which in turn promotes fouling prevention
and facilitates improved separation rates (Muthukumaran et al., 2004; Qiao et al., 2007).
9.5 CONCLUSION
Ultrasound-assisted processing, a novel technology for non-thermal applications, has been

utilized as an alternative to conventional thermal processing. Higher temperature and pressure gen-
erated by high-intensity ultrasound during processing provide a favourable environment for a number
of chemical reactions and microbial inactivation. High-intensity ultrasound generates strong shear
stress, turbulence, and agitation, which help in mixing during food processing. Use of analytical
ultrasound in the food industry will have many benefits such as non-contact and non-destructive
monitoring, economical implementation, rapid measurements, and accurate evaluation. Therefore,
research in the area of analytical ultrasound would help in the development of tools for real-time
monitoring of food processing operations in the future. Destruction of microorganisms and inactiva-
tion of enzymes at low or moderate temperatures in combination with other hurdles without changing
organoleptic and nutritional properties show that the ultrasound-based technologies may have a high
potential for the development of new generation value-added food products. Although ultrasonics
may not completely replace traditional processing, it can be used to complement them to improve
the quality of food products at economical cost. Integration of ultrasonics with other unit operations
such as extraction, cutting, blanching, drying, rehydration, frying, freezing, and thawing can be used
to improve process efficiency. To date, ultrasonic frequency available commercially is limited, and
use of different frequencies along with varying parameters such as temperature, treatment time, and
acoustic power needs further study.
rEFErENCES
Aadil, R.M., Zeng, X.A., Zhang, Z.H., Wang, M.S., Han, Z., Jing, H., and Jabbar, S. (2015). Thermosonication:
A potential technique that influences the quality of grapefruit juice. Int J Food Sci Technol. 50:
1275–1282.
Abid, M., Jabbar, S., Hu, B., Hashim, M.M., Wu, T., Lei, S., Khan, M.A., and Zeng, X. (2014). Thermosonication
as a potential quality enhancement technique of apple juice. Ultrason Sonochem. 21: 984–990.
Acton, E., and Morris, G.J. (1992). Method and apparatus for the control of solidification in liquids. WO. 99:
20420. USA patent application.
Adekunte, A.O., Tiwari, B.K., Cullen, P.J., Scannell, A.G.M., and O’Donnell, C.P. (2010b). Effect of sonica-
tion on colour, ascorbic acid and yeast inactivation in tomato juice. Food Chem. 122: 500–507.
Adekunte, A., Tiwari, B.K., Scannell, A., Cullen, P.J., and O’Donnell, C.P. (2010c). Modelling of yeast inacti-
vation in sonicated tomato juice. Int J Food Microbiol. 137: 116–120.
Adekunte, A., Valdramidis, V.P., Tiwari, B.K., Slone, N., Cullen, P.J., Donnell, C.P.O., and Scannell, A. (2010a).
Resistance of Cronobacter sakazakii in reconstituted powdered infant formula during ultrasound at con-
trolled temperatures: A quantitative approach on microbial responses. Int J Food Microbiol. 142: 53–59.
Afari, G., Yen-Con, H., and King, C.H. (2015). Combination of ultrasound and near neutral electrolyzed
(NEO) water in reducing Escherichia coli O157:H7 and Salmonella Typhimurium DT 104 inoculated
on romaine lettuce leaves. J Food Protect. 78: 281–282.
Alarcon-Rojo, A.D., Janacua, H., Rodriguez, J.C., Paniwnyk, L., and Mason, T.J. (2015). Power ultrasound in
meat processing. Meat Sci. 107: 86–93.
Alves, L.D.L., Cichoski, A.J., Barin, J.S., Rampelotto, C., and Durante, E.C. (2013). The ultrasound on meat
tenderization. Ciencia Rural. 43: 1522–1528.
Anaya-Esparza, L.M., Velazquez-Estrada, R.M., Roig, A.X., Garcia-Galindo, H.S., Sayago-Ayerdi, S.G., and
Montalvo-Gonzalez, E. (2017). Thermosonication: An alternative processing for fruit and vegetable
juices. Trends Food Sci Technol. 61: 26–37.
Ansaria, A.J., Ismaila, M., and Farid, M. (2017). Investigation of the use of ultrasonication followed by heat
for spore inactivation. Food Bioproducts Process. 104: 32–39.
Arroyo, C., Cebrian, G., Pagan, R., and Condon, S. (2012). Synergistic combination of heat and ultrasonic
waves under pressure for Cronobacter sakazakii inactivation in apple juice. Food Control. 25: 342–348.
Arzeni, C., Martinez, K., Zema, P., Arias, A., Perez, O.E., and Pilosof, A.M.R. (2012). Comparative study of
high intensity ultrasound effects on food proteins functionality. J Food Eng. 108: 463–472.
Azoubel, P.M., Baima, M.D.A.M., Amorim, M.D.R., and Oliveira, S.S.B. (2010). Effect of ultrasound on
banana cv Pacovan drying kinetics. J Food Eng. 97: 194–198.
Bashari, M., Zhengyu, J., Jinpeng, W., and Xiaobei, Z. (2016). A novel technique to improve the biodegradation
efficiency of dextranase enzyme using the synergistic effects of ultrasound combined with microwave
shock. Innov Food Sci Emerg Technol. 35: 125–132.
Bayar, N., Bouallegue, T., Achour, M., Kriaa, M., Bougatef, A., and Kammoun, R. (2017). Ultrasonic extrac-
tion of pectin from Opuntia ficus indica cladodes after mucilage removal: Optimization of experimental
conditions and evaluation of chemical and functional properties. Food Chem. 235: 275–282.
Bermudez-Aguirre, D., and Barbosa-Canovas, G.V. (2012). Inactivation of Saccharomyces cerevisiae in pine-
apple, grape and cranberry juices under pulsed and continuous thermo-sonication treatments. J Food
Eng. 108: 383–392.
Benedito, J., Carcel, J.A., Sanjuan, N., and Mulet, A. (2000). Use of ultrasound to assess cheddar cheese
characteristics. Ultrasonics. 38: 727–730.
Bevilacqua, A., Sinigaglia, M., and Corbo, M.R. (2013). Ultrasound and antimicrobial compounds: A suitable
way to control Fusarium oxysporum in juices. Food Bioprocess Technol. 6: 1153–1163.
Butz, P., and Tauscher, B. (2002). Emerging technologies: chemical aspects. Food Res Int. 35: 279–284.
Cappelletti, M., Ferrentino, G., and Spilimbergo, S. (2014). Supercritical carbon dioxide combined with high
power ultrasound: An effective method for the pasteurization of coconut water. J Supercrit Fluids. 92:
257–263.
Cassani, L., Tomadoni, B., Ponce, A., Aguero, M.V., and Moreira, M.R. (2017). Combined use of ultrasound
and vanillin to improve quality parameters and safety of strawberry juice enriched with prebiotic fibers.
Food Bioprocess Technol. 10: 1454–1465.
Cavuldak, O.A., Vural, N., and Anli, R.E. (2016). Ultrasound assisted extraction of plant-derived phenolic
compounds. Gida. 41: 53–60.
Chanukya, B.S., and Rastogi, N.K. (2017). Ultrasound assisted forward osmosis concentration of fruit juice
and natural colorant, Ultrasonics Sonochem. 34: 426–435.
Char, C.D., Mitilinaki, E., Guerrero, S.N., and Alzamora, S.M. (2010). Use of high-intensity ultrasound and
UV-C light to inactivate some microorganisms in fruit juices. Food Bioprocess Technol. 3: 797–803.
Chemat, F., Rombaut, N., Meullemiestre, A., Turk, M., Perino, S., Fabiano-Tixier, A.S., and Abert-Vian, M.
(2017). Review of green food processing techniques. Preservation, transformation, and extraction. Innov
Cheng, L., Da-Wen, S., Zhu, Z., and Zhang, Z. (2017). Emerging techniques for assisting and accelerating
food freezing processes: A review of recent research progresses. Crit Rev Food Sci Nutr. 57: 769–781.
Cheng, X.F., Kai-li, J., Yu-gang, Z., Hua, H., and Min, Z. (2016). The mechanism of enzyme inactivation
induced by ultrasound and its application in food processing. Sci Technol Food Ind. 8: 351–357.
Cheng, X.F., Zhang, M., and Adhikari, B. (2014). Effect of ultrasonically induced nucleation on the drying
kinetics and physical properties of freeze-dried strawberry. Drying Technol. 32: 1857–1864.
Cho, B.K., and Irudayaraj, J.M.K. (2003). Foreign object and internal disorder detection in food materials
using noncontact ultrasound imaging. J Food Sci. 68(3): 967–974.
Chow, R., Blindt, R., Chivers, R., and Povey, M. (2003). The sonocrystallisation of ice in sucrose solutions:
Primary and secondary nucleation. Ultrasonics. 41: 595–604.
Chow, R., Blind, R., Chivers, R., and Povey, M. (2005). A study on the primary and secondary nucleation of
ice by power ultrasound. Ultrasonics. 43: 227–230.
Chow, R., Blind, R., Kamp, A., Grocutt, P., and Chivers, R. (2004). The microscopic visualisation of the sono-
crystallisation of ice using a novel ultrasonic cold stage. Ultrason Sonochem. 11: 245–250.
Clodoveo, M.L., Durante, V., Notte, D. la., Punzi, R., and Gambacorta, G. (2013). Ultrasound-assisted extrac-
tion of virgin olive oil to improve the process efficiency. Eur J Lipid Sci Technol. 115: 1062–1069.
Comandini, P., Blanda, G., Soto-Caballero, M.C., Sala, V., Tylewicz, U., Mujica-Paz, H., Valdez Fragoso,
A., and Gallina Toschi, T. (2013). Effects of power ultrasound on immersion freezing parameters of
potatoes. Innov Food Sci Emerg Technol. 18, 120–125.
Condon, S., Raso, J., and Pagan, R. (2004). Microbial inactivation by ultrasound. In: Novel Food Processing
Technologies, pp. 423–442. Barbosa-Canovas, G.V., Tapia, M.S., and Pilar-Cano, M. Eds., CRC Press,
New York.
Condon-Abanto, S., Arroyo, C., Alvarez, I., Condon, S., Lyng, J.G. (2016). Application of ultrasound in com-
bination with heat and pressure for the inactivation of spore forming bacteria isolated from edible crab
(Cancer pagurus). Int J Food Microbiol. 223: 9–16.
Correia, L.R., Mittal, G.S., and Basir, O.A. (2008). Ultrasonic detection of bone fragment in mechanically
deboned chicken breasts. Innov. Food Sci. Emerg. Technol. 9: 109–115.
Costa, M.G.M., Fonteles, T.V., Jesus, A.L.T. de., Almeida, F.D.L., Miranda, M.R.A.de., Fernandes, F.A.N.,
and Rodrigues, S. (2013). High-intensity ultrasound processing of pineapple juice. Food Bioprocess
Technol. 6: 997–1006.
Cruz, R.M.S., Vieira, M.C., and Silva, C.L.M. (2006). Effect of heat and thermosonication treatments on per-
oxidase inactivation kinetics in watercress (Nasturtium officinale). J Food Eng. 72: 8–15.
Cruz-Cansino, N.del.S., Montiel-Columna, N.I., Bautista-Velueta, P.G., Perez-Tinoco, M.R., Alanis-Garcia, E., and
Ramirez-Moreno, E. (2016). Optimization of thermoultrasound conditions for the processing of a prickly
pear juice blend (Opuntia ficus indica) using response surface methodology. J Food Qual. 39: 780–791.
Daus, A., Horev, B., Dvir, O., Ish-Shalom, S., and Lichter, A. (2011). The efficacy of ultrasonic fumigation for
disinfestation of storage facilities against postharvest pathogens. Postharvest Biol Technol. 62: 310–313.
Delgado, A.E., and Sun, D.W. (2011). Ultrasound-assisted freezing. In: Ultrasound Technologies for Food and
Bioprocessing, pp. 495–509. Feng, H., Barbosa-Canovas, G.V., and Weiss, J. Eds., Springer, New York.
Delgado, A.E., Zheng, L., and Sun, D.W. (2009). Influence of ultrasound on freezing rate of immersion-frozen
apples. Food Bioprocess Technol. 2: 263–270.
Demirdoven, A., and Baysal, T. (2009). The use of ultrasound and combined technologies in food preserva-
tion. Food Rev Int. 25: 1–11.
Demirdoven, A., Ozdogan, K., and Erdogan-Tokatli, K. (2015). Extraction of anthocyanins from red cabbage
by ultrasonic and conventional methods: Optimization and evaluation. J Food Biochem. 39: 491–500.
Dodds, J., Espitalier, F., Louisnard, O., Grossier, R., David, R., Hassoun, M., Baillon, F., Gatumel, C., and
Lyczko, N. (2007). The effect of ultrasound on crystallisation-precipitation processes: Some examples
and a new segregation model. Par Syst Char. 24: 18–28.
Dou, K.N., Yin-de, L., Yu-lan, L., and Cun-guo, G. (2016). Optimization of ultrasonic-assisted extraction
process of essential oils from Glycyrrhiza by response surface methodology. China Cond. 41: 109–112.
Engmann, F.N., Yongkun, M., Tchabo, W., and Hui, M. (2015). Ultrasonication treatment effect on antho-
cyanins, color, microorganisms and enzyme inactivation of mulberry (Moraceae nigra) juice. J Food
Espinosa-Munoz, V., Roldan-Cruz, C.A., Hernandez-Fuentes, A.D., Quintero-Lira, A., Almaraz-Buendia, I.,
and Campos-Montiel, R.G. (2017). Ultrasonic-assisted extraction of phenols, flavonoids, and biocom-
pounds with inhibitory effect against Salmonella typhimurium and Staphylococcus aureus from cactus
pear. J Food Process Eng. 40: e12358.
Evelyn, E., Kim, H.J., and Silva, F.V.M. (2016). Modelling the inactivation of Neosartorya fischeri ascospores
in apple juice by high pressure, power ultrasound and thermal processing. Food Control. 59: 530–537.
Evelyn, E., and Silva, F.V.M. (2015). Use of power ultrasound to enhance the thermal inactivation of Clostridium
perfringens spores in beef slurry. Int J Food Microbiol. 206: 17–23.
Evelyn, E., and Silva, F.V.M. (2016). High pressure processing pretreatment enhanced the thermosonication
inactivation of Alicyclobacillus acidoterrestris spores in orange juice. Food Control. 62: 365–372.
Farahmand, A., Varidi, M., and Koocheki, A. (2016). Investigation of functional properties of quince seed
mucilage extracted by ultrasound. Iranian Food Sci Technol Res J. 12: 163–181.
Fellows, P. (2000). Food Processing Technology: Principles and Practice. CRC Press, New York.
Feng, H., and Yang, W. (2005). Power ultrasound. In: Handbook of Food Science, Technology and Engineering,
pp. 121–127. Hui, Y.H., Ed., CRC Press, New York.
Feng, T., Jing-xin, S., Xin-tao, X., Mei-ling, Y., and Xing-lian, X. (2014). Comparisons of standing, tumbling
and ultrasound treatment on curing of fresh chicken. Food Sci Technol. 39: 111–116.
Fengwei, Y., Kai, F., Jie, H., and Mengxiang, G. (2015). Ultrasonic-assisted solvent extraction of carotenoids
from rapeseed meal: Optimization using response surface methodology. J Food Qual. 38: 377–386.
Fernandes, F.A.N., Gallao, M.I., and Rodrigues, S. (2009). Effect of osmosis and ultrasound on pineapple cell
tissue structure during dehydration. J Food Eng. 90: 186–190.
Fernandez-Ronco, M.P., Gracia, I., Lucas, A. de, and Rodriguez, J.F. (2013). Extraction of Capsicum annuum
oleoresin by maceration and ultrasound-assisted extraction: Influence of parameters and process
modeling. J Food Process Eng. 36: 343–352.
Ferrante, S., Guerrero, S., and Maris Alzamora, S. (2007). Combined use of ultrasound and natural antimicro-
bials to inactivate Listeria monocytogenes in orange juice. J Food Protect. 70: 1850–1856.
Ferrario, M., Alzamora, S.M., and Guerrero, S. (2015). Study of the inactivation of spoilage microorganisms
in apple juice by pulsed light and ultrasound. Food Microbiol. 46: 635–642.
Fijalkowska, A., Nowacka, M., Wiktor, A., Sledz, M., and Witrowa-Rajchert, D. (2016). Ultrasound as a pre-
treatment method to improve drying kinetics and sensory properties of dried apple. J Food Process Eng.
39: 256–265.
Filho, E.G.A., Cullen, P.J., Frias, J.M., Bourke, P., Tiwari, B.K., Brito, E.S., Rodrigues, S., and Fernandes,
F.A.N. (2016). Evaluation of plasma, high-pressure and ultrasound processing on the stability of fruc-
tooligosaccharides. Int J Food Sci Technol. 51: 2034–2040.
Fonteles, T.V., Costa, M.G.M., De Jesus, A.L.T., de Miranda, A.R.M., Fernandes, F.A.N., and Rodrigues, S.
(2012). Power ultrasound processing of cantaloupe melon juice: Effects on quality parameters. Food Res
Int. 48: 41–48.
Gallego-Juarez, J.A., Riera, E., Fuente Blanco, S.D.L., Rodriguez, C.G., Acosta, A.V.M., and Blanco, A.
(2007). Application of high-power ultrasound for dehydration of vegetables: Processes and devices.
Drying Technol. 25: 1893–1901.
Ganjloo, A., Rahman, R.A., Bakar, J., Osman, A., and Bimakr, M. (2008). Feasibility of high-intensity ultra-
sonic blanching combined with heating for peroxidase inactivation of seedless guava (Psidium guajava
L.). In: Proceedings of the 18th National Congress in Food Technology, Mashhad, Iran, October 15–16.
http://confnews.um.ac.ir/images/41/conferences/foodcongress/pdfs/oe_03.pdf.
Garcia-Perez, J.V., Carcel, J.A., Benedito, J., and Mulet, A. (2007). Power ultrasound mass transfer enhance-
ment in food drying. Food Bioprod Process. 85: 247–254.
Garud, S.R., Priyanka, B.S., Negi, P.S., and Rastogi, N.K. (2017). Effect of thermosonication on bacterial count
in artificially inoculated model system and natural microflora of sugarcane juice. J Food Process Preserv.
41: e12813.
Gayas, B., Kaur, G., and Gul, K. (2017). Ultrasound-assisted extraction of apricot kernel oil: Effects on func-
tional and rheological properties. J Food Process Eng. 40: e12439.
Gomez, M., Oliete, B., Garcalvarez, J., Ronda, F., and Salazar, J. (2008). Characterization of cake batters by
ultrasound measurements J. Food Eng. 89: 408–413.
Grossier, R., Louisnard, O., and Vargas, Y. (2007). Mixture segregation by an inertial cavitation bubble.
Ultrason Sonochem. 14: 431–437.
Hengl, N., Jin, Y., Pignona, F., Baup, S., Mollard, R., Gondrexon, N., Magnin, A., Michotb, L., and Paineauc, E.
(2014). A new way to apply ultrasound in cross-flow ultrafiltration: Application to colloidal suspensions.
Ultrason Sonochem. 21: 1018–1025.
Hiranrangsee, L., Kumaree, K.K., Sadiq, B.M., and Anal, A.K. (2016). Extraction of anthocyanins from pericarp
and lipids from seeds of mangosteen (Garcinia mangostana L.) by ultrasound-assisted extraction (UAE)
and evaluation of pericarp extract enriched functional ice-cream. J Food Sci Technol. 53: 3806–3813.
Hong, G.P., Ji-Yeon, C., Yeon-Ji, J., and Mi-Jung, C. (2014). Effects of water or brine immersion thawing com-
bined with ultrasound on quality attributes of frozen pork loin. Korean J Food Sci Animal Res. 34: 115–121.
Hosseinzadeh, S.B., Hadi-Khoshtaghaza, M., Lorigooini, Z., Minaei, S., and Zareiforoush, H. (2015). Analysis
of the combinative effect of ultrasound and microwave power on Saccharomyces cerevisiae in orange
juice processing. Innov Food Sci Emerg Technol. 32: 110–115.
Hou, F., Dongxiao, S., Jinrui, X., Yushi, G., Zhang, R., Wei, Z., Chi, J., and Zhang, M. (2016). Enhanced
extraction of phenolics and antioxidant capacity from sorghum (Sorghum bicolor L. Moench) shell
using ultrasonic-assisted ethanol-water binary solvent. J Food Process Preserv. 40: 1171–1179.
Hu, H., Hu, T., Xu, Q., Wang, T., Wang, K., Xu, X., and Pan, S. (2015). A review of recent studies on high-intensity
ultrasound in food protein processing. Food Sci China. 36: 260–265.
Hu, S.Q., Liu, G., Li, L., Li, Z.X., and Hou, Y. (2013). An improvement in the immersion freezing process for
frozen dough via ultrasound irradiation. J Food Eng. 114: 22–28.
Islam, M.N., Zhang, M., and Adhikari, B. (2014a). The inactivation of enzymes by ultrasound: A review of
potential mechanisms. Food Rev Int. 30: 1–21.
Islam, M.N., Zhang, M., Adhikari, B., Cheng, X.F., and Xu, B.G. (2014b). The effect of ultrasound-assisted
immersion freezing on selected physicochemical properties of mushrooms. Int J Refrig. 42: 121–133.
Jabbar, S., Abid, M., Hu, B., Hashim, M.M., Lei, S., Wu, T., and Zeng, X. (2015b). Exploring the potential of
thermosonication in carrot juice processing. J Food Sci Technol. 52: 7002–7013.
Jabbar, S., Abid, M., Wu, T., Hashim, M.M., Saeeduddin, M., Hu, B., Lei, S., and Zeng, X. (2015a). Ultrasound-
assisted extraction of bioactive compounds and antioxidants from carrot pomace: A response surface
approach. J Food Process Preserv. 39: 1878–1888.
Jain, S., and Anal, A.K. (2016). Optimization of extraction of functional protein hydrolysates from chicken
egg shell membrane (ESM) by ultrasonic assisted extraction (UAE) and enzymatic hydrolysis. LWT-
Jang, J.H., and Moon, K.D. (2011). Inhibition of polyphenol oxidase and peroxidase activities on fresh-cut
apple by simultaneous treatment of ultrasound and ascorbic acid. Food Chem. 124: 444–449.
Jayani, C., and Leong, T. (2015). Ultrasonic processing for dairy applications: Recent advances. Food Eng
Rev. 7: 143–158.
Jimenez-Sanchez, C., Lozano-Sanchez, J., Segura-Carretero, A., and Fernandez-Gutierrez, A. (2017a).
Alternatives to conventional thermal treatments in fruit-juice processing. Part 1: Techniques and appli-
cations. Crit Rev Food Sci Nutr. 57: 501–523.
Jimenez-Sanchez, C., Lozano-Sanchez, J., Segura-Carretero, A., and Fernandez-Gutierrez, A. (2017b).
Alternatives to conventional thermal treatments in fruit-juice processing. Part 2: Effect on composition,
phytochemical content, and physicochemical, rheological, and organoleptic properties of fruit juices.
Crit Rev Food Sci Nutr. 57: 637–652.
Jin, S., and Ma, K.J. (2017). Research progress in ultrasound-assisted extraction of carotenoids. Food Res Dev.
38: 192–197.
Jose, J.F.B.de.S., Medeiros, H.S.de., Bernardes, P.C., and Andrade, N.J.de. (2014). Removal of Salmonella
enterica Enteritidis and Escherichia coli from green peppers and melons by ultrasound and organic
acids. Int J Food Microbiol. 190: 9–13.
Kadam, S.U., Tiwari, B.K., Álvarez, C., and O’Donnell, C.P. (2015). Ultrasound applications for the extraction,
identification and delivery of food proteins and bioactive peptides. Trends Food Sci. Technol. 46: 60–67.
Kiang, W.S., Bhat, R., Rosma, A., and Cheng, L.H. (2013). Effects of thermosonication on the fate of
Escherichia coli O157: H7 and Salmonella enteritidis in mango juice. Lett Appl Microbiol. 56: 251–257.
Kiani, H., Sun, D.W., and Zhang, Z.H. (2012). The effect of ultrasound irradiation on the convective heat trans-
fer rate during immersion cooling of a stationary sphere. Ultrason Sonochem. 19: 1238–1245.
Knorr, D., Zenker, M., Heinz, V., and Lee, D.U. (2004) Applications and potential of ultrasonics in food
processing. Trends Food Sci. Technol. 15:261–166.
Koshani, R., Ziaee, E., Niakousari, M., and Golmakani, M.T. (2014). Optimization of thermal and thermosoni-
cation treatments on pectin methyl esterase inactivation of sour orange juice (Citrus aurantium). J Food
Kumari, B., Tiwari, B.K., Hossain, M.B., Rai, D.K., and Brunton, N.P. (2017). Ultrasound-assisted extraction of
polyphenols from potato peels: Profiling and kinetic modelling. Int J Food Sci Technol. 52: 1432–1439.
Kyllonen, H.M., Pirkonen, P., and Nystrom, M. (2005). Membrane filtration enhanced by ultrasound: A review.
Desalination. 181: 319–335.
Lei, C., Yan-bin, X., Zai-quan, C., and Mei Ren. (2016). Progress on application of ultrasound treatment tech-
nology in meat industry. Food Machinery. 5: 232–236.
Li, B., and Sun, D.W. (2002). Effect of power ultrasound on freezing rate during immersion freezing. J Food
Eng. 55: 277–282.
Li, H.Z., Pordesimo, L., and Weiss, J. (2004). High intensity ultrasound assisted extraction of oil from soybeans.
Food Res Int. 37: 731–738.
Li, Y., Chun-ran, H., He-yan, H., Chuan-rong, D., Jia-cheng, Z., and Cong-yu, L. (2017). Optimization of
extraction process of total flavonoids from lingonberry and evaluation of its antibacterial activities.
Food Res Dev. 38: 69–74.
Lin, S.X.Q., and Chen, X.D. (2007). A laboratory investigation of milk fouling under the influence of ultra-
sound. Food Bioprod. Process. 85: 57–62.
Lin, Z., Zhang, Z., Chen, X., and Ming, Q. (2011). Ultrasonic-assisted extraction of red rice pigment. Modern
Liu, L., Jia, W., Xu, D., and Li, R. (2015). Applications of ultrasonic cutting in food processing. J Food Process
Preserv. 39: 1762–1769.
Liu, Y., Jianye, W., Cuijuan, C., and Shuai, M. (2014). Ultrasound assisted osmotic dehydration as pretreatment
for hot-air drying of carrot. Food Sci Technol Res. 20: 31–41.
Lu, H.Q., Xie, C.F., Yang, R.F., and Qiu, T.Q. (2005). Preventing fouling in evaporators through ultrasound.
Int. Sugar J. 107: 456–461.
Luo, K., and Oh, D.H. (2015). Synergistic effect of slightly acidic electrolyzed water and ultrasound at mild
heat temperature in microbial reduction and shelf-life extension of fresh-cut bell pepper. J Microbiol
Biotechnol. 25: 1502–1509.
Luo, K., Kim, S.Y., Wang, J., and Oh, D.H. (2016). A combined hurdle approach of slightly acidic electrolyzed
water simultaneous with ultrasound to inactivate Bacillus cereus on potato. LWT-Food Sci Technol. 73:
615–621.
Luo, K., and Oh, D.H. (2016). Inactivation kinetics of Listeria monocytogenes and Salmonella enterica
serovar Typhimurium on fresh-cut bell pepper treated with slightly acidic electrolyzed water combined
with ultrasound and mild heat. Food Microbiol. 53: 165–171.
Ma, K.J., Si, J., and Yan-liang, P. (2016). Research status and prospect of ultrasonic technology in food devel-
opment. Food Ind. 9: 207–211.
Marx, G., Moody, A., and Bermudez-Aguirre, D. (2011). A comparative study on the structure of Saccharomyces
cerevisiae under nonthermal technologies: High hydrostatic pressure, pulsed electric fields and thermo-
sonication. Int J Food Microbiol. 151: 327–337.
Mason, T.J. (1998). Power ultrasound in food processing–the way forward. In: Ultrasound in Food Processing,
pp. 105–126. Povey, M.J.W., and Mason, T.J., Eds., Thomson Science, London, UK.
Mason, T.J. (1999). Sonochemistry. Oxford Science Publications, Oxford, MS.
Mason, T.J., and Peters, D. (2002). Practical Sonochemistry, Power Ultrasound Uses and Applications. Ellis
Horwood Publishers, Chichester, UK.
Mason, T.J., Riera, E., Vercet, A., and Buesa, P.L. (2005). Application of ultrasound. In: Emerging Technologies
for Food Processing, pp. 323–332. Sun, D.W. Ed., Elsevier, London, UK.
McClements, D.J. (1995). Advances in the application of ultrasound in food analysis and processing. Trends
McDonnell, C.K., Lyng, J.G., and Allen, P. (2014). The use of power ultrasound for accelerating the curing of
pork. Meat Sci. 98: 142–149.
Mead, E.L., Sutherland, R.G., and Verrall, R.E. (1976). The effect of ultrasound on water in the presence of
dissolved gases. Can J Chem. 54: 1114–1120.
Melgar-Lalanne, G., Hernandez-Alvarez, A.J., Jimenez-Fernandez, M., and Azuara, E. (2017). Oleoresins
from Capsicum spp.: Extraction methods and bioactivity. Food Bioprocess Technol. 10: 51–76.
Mello, B.T.F., Garcia, V.A.S., and Silva, C.da. (2017). Ultrasound-assisted extraction of oil from chia (Salvia
hispanica L.) seeds: Optimization extraction and fatty acid profile. J Food Process Eng. 40: e12298.
Meng, X.M., and Ming-hua, L. (2017). Study on extraction technique for red pigments from red pepper.
Storage Process. 17: 68–72.
Miks-Krajnik, M., Feng, J., Woo Suk Bang, L.X., and Hyun-Gyun, Y. (2017). Inactivation of Listeria mono-
cytogenes and natural microbiota on raw salmon fillets using acidic electrolyzed water, ultraviolet light
or/and ultrasounds. Food Control. 74: 54–60.
Mohammadi, V., Ghasemi-Varnamkhasti, M., and Gonzalez, L.A. (2017). Analytical measurements of ultra-
sound propagation in dairy products: A review. Trends Food Sci Technol. 61: 38–48.
Mongenot, N., Charrier, S., and Chalier, P. (2000). Effect of ultrasound emulsification on cheese aroma encap-
sulation by carbohydrates. J. Agric. Food Chem. 48: 861–867.
Morsy, N.F.S. (2016). A comparative study of nutmeg (Myristica fragrans Houtt.) oleoresins obtained by con-
ventional and green extraction techniques. J Food Sci Technol. 53: 3770–3777.
Moser, W.R., Find, J., Emerson, S.C., and Krausz, I.M. (2001). Engineered synthesis of nanostructured materi-
als and catalysts. Adv Chem Eng. 27: 1–48.
Mugdha, P.D., and Ananthanarayan, L. (2017). Comparative inactivation studies of Aegle marmelos (bael)
peroxidase in crude extract of fruit by heat processing and ultrasonication treatment. J Food Meas Char.
11: 417–422.
Munoz, A., Caminiti, I.M., Palgan, I., Pataro, G., Noci, F., Morgan, D.J., et al. (2012). Effects on Escherichia
coli inactivation and quality attributes in apple juice treated by combinations of pulsed light and ther-
mosonication. Food Res Int. 45: 299–305.
Munoz, A., Palgan, I., Noci, F., Morgan, D.J., Cronin, D.A., Whyte, P., and Lyng, J.G. (2011). Combinations of
high intensity light pulses and thermosonication for the inactivation of Escherichia coli in orange juice.
Food Microbiol. 28: 1200–1204.
Musielak, G., Mierzwa, D., and Kroehnke, J. (2016). Food drying enhancement by ultrasound: A review.
Trends Food Sci Technol. 56: 126–141.
Muthukumaran, S., Yang, K., Seuren, A., Kentish, S., Ashokkumar, M., Stevens, G.W., and Grieser F. (2004).
The use of ultrasonic cleaning for ultrafiltration membranes in the dairy industry. Sep. Purif. Technol.
39: 99–107.
Muzaffar, S., Ahmad, M., Wani, S.M., Adil, G., Waqas, N.B., Shah, U., et al. (2016). Ultrasound treatment:
Effect on physicochemical, microbial and antioxidant properties of cherry (Prunus avium). J Food Sci
Technol. 53: 2752–2759.
Narayan, A.V., Nagaraj, N., Hebbar, H.U., Chakkaravarthi, A., Raghavarao, K.S.M.S., and Nene, S. (2000).
Acoustic field-assisted osmotic membrane distillation. Desal. 147: 149–156.
Nour, V., Trandafir, I., and Cosmulescu, S. (2016). Optimization of ultrasound-assisted hydroalcoholic extraction of
phenolic compounds from walnut leaves using response surface methodology. Pharm Biol. 54: 2176–2187.
Nowacka, M., Wiktor, A., Sledz, M., Jurek, N., and Witrowa-Rajchert, D. (2012). Drying of ultrasound pre-
treated apple and its selected physical properties. J Food Eng. 113: 427–433.
O’Sullivan, J., Arellano, M., Pichot, R., and Norton, I.T. (2014). The effect of ultrasound treatment on the
structural, physical and emulsifying properties of dairy proteins. Food Hydrocol. 42: 386–396.
O’Sullivan, J.J., Park, M., Beevers, J., Greenwood, R.W., and Norton, I.T. (2017). Applications of ultrasound for the
functional modification of proteins and nanoemulsion formation: A review. Food Hydrocol. 71: 299–310.
Ozcan, G., and Zorba, N.N.D. (2016). Combined effect of ultrasound and essential oils to reduce Listeria
monocytogenes on fresh produce. Food Sci Technol Int. 22: 353–362.
Ozuna, C., Paniagua-Martinez, I., Castano-Tostado, E., Ozimek, L., and Amaya-Llano, S.L. (2015). Innovative
applications of high-intensity ultrasound in the development of functional food ingredients: Production
of protein hydrolysates and bioactive peptides. Food Res Int. 77: 685–696.
Pakrokh Ghavi, P. (2015). Modeling and optimization of ultrasound-assisted extraction of polysaccharide

from the roots of Althaea officinalis. J Food Process Preserv. 39: 2107–2118.
Pang, X.Y., Xiu-fang, X., and Bao-hua, K. (2017). The mechanism of ultrasonic assisted refrigeration technol-
ogy and its application in frozen food. Food Res Dev. 38: 190–194.
Park, S.Y., Hyun-Ha, S., and Sang-Do, H.A. (2014). Synergistic effects of NaOCl and ultrasound combina-
tion on the reduction of Escherichia coli and Bacillus cereus in raw laver. Foodborne Pathog Dis. 11:
373–378.
Park, S.Y., Rahaman Mizan, M.F., Ha, A., Yang, S.D., and Sang-Do, H.A. (2015). Inactivation of Cronobacter
sakazakii in head lettuce by using a combination of ultrasound and sodium hypochlorite. J Food Prot.
78: 249.
Patil, S., Bourke, P., Kelly, B., Frias, J.M., and Cullen, P.J. (2009). The effects of acid adaptation on Escherichia
coli inactivation using power ultrasound. Innov Food Sci Emerg Technol. 10: 486–490.
Piyasena, P., Mohareb, E., and McKellar, R.C. (2003). Inactivation of microbes using ultrasound: A review.
Int J Food Microbiol. 87: 207–216.
Qiao, X.L., Zhang, Z.J., and Ping, Z.H. (2007). Hydrophilic modification of ultrafiltration membranes and
their application in Salvia miltiorrhiza decoction. Sep. Purif. Technol. 56: 265–269.
Rabelo, R.S., Machado, M.T.C., Martinez, J., and Hubinger, M.D. (2016). Ultrasound assisted extraction and
nanofiltration of phenolic compounds from artichoke solid wastes. J Food Eng. 178: 170–180.
Rastogi, N.K. (2011). Opportunities and challenges in application of ultrasound in food processing. Crit Rev
Food Sci Nutr. 51: 705–722.
Ravanfa, R., Mohammad Tamadon, A., and Mehrdad, N. (2015). Optimization of ultrasound assisted extrac-
tion of anthocyanins from red cabbage using Taguchi design method. J Food Sci Technol. 52: 8140–8147.
Raviyan, P., Zhang, Z., and Feng, H. (2005). Ultrasonication for tomato pectinmethylesterase inactivation:
Effect of cavitation intensity and temperature on inactivation. J Food Eng. 70: 189–196.
Raza, A., Li, F., Xu, X., and Tang, J. (2017). Optimization of ultrasonic-assisted extraction of antioxidant
polysaccharides from the stem of Trapa quadrispinosa using response surface methodology. Int J Biol
Macromol. 94: 335–344.
Ribeiro, M.de.M., Valdramidis, V.P., Nunes, C.A., and Souza, V.R.de. (2017). Synergistic effect of thermo-
sonication to reduce enzymatic activity in coconut water. Innov Food Sci Emerg Technol. 41: 404–410.
Rodgers, S.L., and Ryser, E.T. (2004). Reduction of microbial pathogens during apple cider production using
sodium hypochlorite, copper ion, and sonication. J Food Prot. 67: 766–771.
Rojas, M.L., Leite, T.S., Cristianini, M., Alvim, I.D., and Augusto, P.E.D. (2016). Peach juice processed by
the ultrasound technology: Changes in its microstructure improve its physical properties and stability.
Food Res Int. 82: 22–33.
Rosario, D.K.A. do., Mutz, Y.da.S., Peixoto, J.M.C., Oliveira, S.B.S., Carvalho, R.V.da., Carneiro, J.C.S., et al.
(2017). Ultrasound improves chemical reduction of natural contaminant microbiota and Salmonella
enterica subsp. enterica on strawberries. Int J Food Microbiol. 241: 23–29.
Ruecroft, G., Hipkiss, D., Ly, T., Maxted, N., and Cains, P.W. (2005). Sonocrystallization: The use of ultra-
sound for improved industrial crystallization. Org Process Res Dev. 9: 923–932.
Russell, A.B., Cheney, P.E., and Wantling, S.D. (1998). Influence of freezing conditions on ice crystallisation
in ice cream. J Food Eng. 39: 179–191.
Saclier, M., Peczalski, R., and Andrieu, J. (2010). A theoretical model for ice primary nucleation induced by
acoustic cavitation. Ultrason Sonochem. 17: 98–105.
Sala, F.J., Burgos, J., Condon, S., Lopez, P., and Raso, J. (1995). Effect of heat and ultrasound on microor-
ganisms and enzymes. In: New Methods of Food Preservation, pp. 176–204. Gould, G.W. Ed., Blackie
Academic & Professional, London, UK.
Saleem, R., and Ahmad, R. (2016). Effect of ultrasonication on secondary structure and heat induced gelation
of chicken myofibrils. J Food Sci Technol. 53: 3340–3348.
Sanchez-Rubio, M., Taboada-Rodriguez, A., Cava-Roda, R., Lopez-Gomez, A., and Marin-Iniesta, F. (2016).
Combined use of thermo-ultrasound and cinnamon leaf essential oil to inactivate Saccharomyces cere-
visiae in natural orange and pomegranate juices. LWT-Food Sci Technol. 73: 140–146.
Scheba, G., Weige, R.B., and O’Brien, J.R. (1991). Quantitative assessment of germicidal efficiency of ultra-
sonic energy. Appl Environ Microbiol. 57: 2079–2084.
Schossler, K., Jaeger, H., and Knorr, D. (2012). Novel contact ultrasound system for the accelerated
freeze-drying of vegetables. Innov Food Sci Emerg Technol. 16: 113–120.
Sharmila, G., Nikitha, V.S., Ilaiyarasi, S., Dhivya, K., Rajasekar, V., Manoj Kumar, N., et al. (2016). Ultrasound
assisted extraction of total phenolics from Cassia auriculata leaves and evaluation of its antioxidant
activities. Ind Crops Prod. 84: 13–21.
Silva, L.C.A., Almeida, P.S., Rodrigues, S., and Fernandes, F.A.N. (2015). Inactivation of polyphenoloxidase
and peroxidase in apple cubes and in apple juice subjected to high intensity power ultrasound processing.
J Food Process Preserv. 39: 2081–2087.
Simon, A., Penpenic, L., Gondrexon, N., Taha, S., and Dorange, G. (2000). A comparative study between
classical stirred and ultrasonically assisted dead end ultrafiltration. Ultrasonics Sonochem. 7: 183–186.
Singh, Y., Mazumder, A., Giri, A., and Mishra, H.N. (2017). Optimization of ultrasound-assisted extraction of
beta-carotene from Chlorella biomass (MCC7) and its use in fortification of apple jam. J Food Process
Eng. 40: e12321.
Siucinska, K., Mieszczakowska-Frac, M., Polubok, A., and Konopacka, D. (2016). Effects of ultrasound assis-
tance on dehydration processes and bioactive component retention of osmo-dried sour cherries. J Food
Sci. 81: C1654–C1661.
Stefanovic, A., Jovanovic, J., Dojcinovic, M., Levic, S., Zuza, M., Nedovic, V., and Knežević-Jugović, Z.
(2014a). Impact of high-intensity ultrasound probe on the functionality of egg white proteins. J Hyg Eng
Des. 6: 215–224.
Stefanovic, A.B., Jovanovic, J.R., Grbavcic, S.Z., Sekuljica, N.Z., Manojlovic, V.B., Bugarski, B.M., and
Knežević-Jugović, Z.D. (2014b). Impact of ultrasound on egg white proteins as a pretreatment for func-
tional hydrolysates production. European Food Res Technol. 239: 979–993.
Sun, D.W., and Li, B. (2003). Microstructural change of potato tissues frozen by ultrasound-assisted immer-
sion freezing. J Food Eng. 57: 337–345.
Sun, D.W., and Zheng, L.Y. (2006). Innovations in freezing process. In: Handbook of Frozen Food Processing
and Packaging, pp. 175–195. Sun, D.W. Ed., The Chemical Rubber Company Press, Boca Raton, FL.
Sun, Y., Hailing, Y., Xueming, Z., Ling, Z., and Wang, W. (2011). Ultrasonic-assisted enzymatic degradation
of cholesterol in egg yolk. Innov Food Sci Emerg Technol. 12: 505–508.
Sureeporn, K., Murkovic, M., and Thongraung, C. (2014). Antioxidant and nitric oxide inhibitory activities
of tilapia (Oreochromis niloticus) protein hydrolysate: Effect of ultrasonic pretreatment and ultrasonic-
assisted enzymatic hydrolysis. Int J Food Sci Technol. 49: 1932–1938.
Tan, M.C., Chin, N.L., and Yusof, Y.A. (2011). Power ultrasound aided batter mixing for sponge cake batter.
J Food Eng. 104: 430–437.
Tenga, M.Y., Lina, S.H., and Juang, R.S. (2006). Effect of ultrasound on the separation of binary protein mix-
tures by cross-flow ultrafiltration. Desal. 200: 280–282.
Terefe, N.S., Gamage, M., Vilkhu, K., Simons, L., Mawson, R., and Versteeg, C. (2009). The kinetics of
inactivation of pectin methylesterase and polygalacturonase in tomato juice by thermosonication. Food
Chem. 117: 20–27.
Tian, M., Wang, W., and Tu, L. (2015). Optimization of ultrasonic-assisted solvent extraction process for azo-
cyclotin from dried jujube by response surface analysis. Food Sci China. 36: 58–62.
Tiwari, B.K., Muthukumarappan, K., O’Donnell, C.P., and Cullen, P.J. (2009). Inactivation kinetics of pectin
methylesterase and cloud retention in sonicated orange juice. Innov Food Sci Emerg Technol. 10: 166–171.
Tomadoni, B., Moreira, M. del R., Espinosa, J.P., and Ponce, A. (2017). Individual and combined effects of
pomegranate extract and ultrasonic treatments on kiwifruit juice quality parameters. J Food Process
Eng. 40: e12339.
Tremarin, A., Brandao, T.R.S., and Silva, C.L.M. (2017). Application of ultraviolet radiation and ultrasound
treatments for Alicyclobacillus acidoterrestris spores inactivation in apple juice. LWT-Food Sci Technol.
78: 138–142.
Tsukamoto, I., Yim, B., Stavarache, C.E., Furuta, M., Hashiba, K., and Maeda, Y. (2004). Inactivation of
Saccharomyces cerevisiae by ultrasonic irradiation. Ultrason Sonochem. 11: 61–65.
Upadhyay, R., Nachiappan, G., and Mishra, H.N. (2015). Ultrasound-assisted extraction of flavonoids and
phenolic compounds from Ocimum tenuiflorum leaves. Food Sci Biotechnol. 24: 1951–1958.
Vercet, A., Sanchez, C., Burgos, J., Montanes, L., and Lopez Buesa, P. (2002). The effects of manothermo-
sonication on tomato pectic enzymes and tomato paste rheological properties. J Food Eng. 53: 273–278.
Walkling-Ribeiro, M., Noci, F., Riener, J., Cronin, D.A., Lyng, J.G., and Morgan, D.J. (2009). The impact of
thermosonication and pulsed electric fields on Staphylococcus aureus inactivation and selected quality
parameters in orange juice. Food Bioprocess Technol. 2: 422–430.
Wambura, P., and Yang, W. (2011). Influence of power ultrasound on oxidative rancidity of potato chips.
J Food Process Eng. 34: 1046–1052.
Wambura, P., Yang, W., and Wang, Y. (2008). Power ultrasound enhanced one-step soaking and gelatinization
for rough rice parboiling. Int. J Food Eng. 4(4): 1–12.
Wang, J., He, D.L., Muhammad, U., Jin, Z.H., Zhao, H.W., Zhao, X.L., et al. (2016). Ultrasound-assisted
extraction of polysaccharides from Artemisia selengensis Turcz and its antioxidant and anticancer activ-
ities. J Food Sci Technol. 53: 1025–1034.
Wang, J., Hu, X., and Wang, Z. (2010). Kinetics models for the inactivation of Alicyclobacillus acidiphilus
DSM14558T and Alicyclobacillus acidoterrestris DSM 3922T in apple juice by ultrasound. Int J Food
Microbiol. 139: 177–181.
Wang, W., Ma, X., Zou, M., Jiang, P., Hu, W., Li, J., et al. (2015). Effects of ultrasound on spoilage
microorganisms, quality, and antioxidant capacity of postharvest cherry tomatoes. J Food Sci. 80:
C2117–C2126.
Wen, Q.H., Zong-Cai, T., Zhang, L., Wang, H., and Chang, H.X. (2017). Effect of high intensity ultrasound on
the gel and structural properties of Ctenopharyngodon idellus myofibrillar protein. J Food Biochem.
41: e12288.
Wu, H., Junxiang, Z., Wenchao, D., and Wang, C. (2014). Ultrasound-assisted enzymatic extraction and
antioxidant activity of polysaccharides from pumpkin (Cucurbita moschata). Carbohydr Pol. 113:
314–324.
Wu, J., Gamage, T.V., Vilkhu, K.S., Simons, L.K., and Mawson, R. (2008). Effect of thermosonication on qual-
ity improvement of tomato juice. Innov Food Sci Emerg Technol. 9: 186–195.
Wu, S.X., Zhou, H.Y., Xia, Y.Y., Shang, Y.B., and Li, H.J. (2012). Methods of improving meat tenderness and
the mechanism of tenderization. Food Sci. Technol. 37(9): 126–129.
Wu, X., and Narsimhan, G. (2017). Synergistic effect of low power ultrasonication on antimicrobial activity of
melittin against Listeria monocytogenes. LWT-Food Sci Technol. 75: 578–581.
Xin, Y., Zhang, M., and Adhikari, B. (2014a). Freezing characteristics and storage stability of broccoli
(Brassica oleracea L. var. botrytis L.) under osmodehydrofreezing and ultrasound-assisted osmodehy-
drofreezing treatments. Food Bioprocess Technol. 7: 1736–1744.
Xin, Y., Zhang, M., and Benu, A. (2014b). The effects of ultrasound-assisted freezing on the freezing time
and quality of broccoli (Brassica oleracea L. var. botrytis L.) during immersion freezing. Int J Refrig.
41: 82–91.
Xu, B., Zhang, M., Bhandari, B., and Cheng, X. (2014a). Influence of power ultrasound on ice nucleation of
radish cylinders during ultrasound-assisted immersion freezing. Int J Refrig. 46: 1–8.
Xu, B., Zhang, M., Bhandari, B., and Cheng, X. (2014b). Influence of ultrasound-assisted osmotic dehydration
and freezing on the water state, cell structure, and quality of radish (Raphanus sativus L.) cylinders.
Drying Technol. 32: 1803–1811.
Xu, Y., Zhang, L., Bailina, Y., Ge, Z., Ding, T., Ye, X., and Liu, D. (2014c). Effects of ultrasound and/or heating
on the extraction of pectin from grapefruit peel. J Food Eng. 126: 72–81.
Xu, B., Zhang, M., Bhandari, B., Cheng, X., and Sun, J. (2015). Effect of ultrasound immersion freezing
on the quality attributes and water distributions of wrapped red radish. Food Bioprocess Technol. 8:
1366–1376.
Yildiz, G., Rababah, T.M., and Feng, H. (2016). Ultrasound-assisted cutting of cheddar, mozzarella and Swiss
cheeses: Effects on quality attributes during storage. Innov Food Sci Emerg Technol. 37: 1–9.
Yolmeh, M., and Najafzadeh, M. (2014). Optimisation and modelling green bean’s ultrasound blanching. Int J
Youssouf, L., Lallemand, L., Giraud, P., Soule, F., Bhaw-Luximon, A., Meilhac, O., et al. (2017). Ultrasound-
assisted extraction and structural characterization by NMR of alginates and carrageenans from sea-
weeds. Carbohyd Pol. 166: 55–63.
Yu, H., Seow, Y.X., Ong, P.K.C., and Zhou, W. (2016). Generating Maillard reaction products in a model
system of D-glucose and L-serine by continuous high-intensity ultrasonic processing. Innov Food Sci
Yuan, Y., Hu, Y., Yue, T., Chen, T., and Lo, Y.M. (2009). Effect of ultrasonic treatments on thermoacidophilic
alicyclobacillus acidoterrestris in apple juice. J Food Process Presev. 33: 370–383.
Zhang, C., Yang, Q., Zhang, H., and Wang, C. (2014). Optimization for ultrasonic-assisted on extraction of
Monascus red pigment and yellow pigments. J Food Sci Biotechnol. 33: 805–813.
Zhang, X., Inada, T., and Tezuka, A. (2003). Ultrasonic-induced nucleation of ice in water containing air
bubbles. Ultrason Sonochem. 10: 71–76.
Zhao, B.S., Basir, O.A., and Mittal, G.S. (2003). Detection of metal, glass and plastic pieces in bottled bever-
ages using ultrasound. Food Res. Int. 36(5): 513–521.
Zhao, C.B., Zhou, L.Y., Liu, J.Y., Zhang, Y., Chen, Y., and Wu, F. (2016). Effect of ultrasonic pretreatment
on physicochemical characteristics and rheological properties of soy protein/sugar Maillard reaction
products. J Food Sci Technol. 53: 2342–2351.
Zhao, L., Zhang, S., Uluko, H., Liu, L., Lu, J., Xue, H., et al. (2014). Effect of ultrasound pretreatment on
rennet-induced coagulation properties of goat’s milk. Food Chem. 165: 167–174.
Zheng, L., and Sun, D.W. (2005). Ultrasonic assistance of food freezing. In: Emerging Technologies for Food
Processing, pp. 603–627. Sun, D.W. Ed., Elsevier, London, UK.
Zheng, L., and Sun, D.W. (2006). Innovative applications of power ultrasound during food freezing processes-
a review. Trends Food Sci Technol. 17: 16–23.
Zhu, C., and Liu, G. (2000). Modeling of ultrasonic enhancement on membrane distillation. J. Membr. Sci.
176: 31–41.
Zinoviadou, K.G., Galanakis, C.M., Brnčić, M., Grimi, N., Boussetta, N., Mota, M.J., Saraiva, J.A., Patras,
A., Tiwari, B.K., and Barba, F.J. (2015). Fruit juice sonication: Implications on food safety and physico-
chemical and nutritional properties. Food Res. Int. 77: 743–752.
Zisu, B., Lee, J., Chandrapala, J., Bhaskaracharya, R., Palmer, M., Kentish, S., and Ashokkumar, M. (2011).
Effect of ultrasound on the physical and functional properties of reconstituted whey protein powders.
J. Dairy Res. 78(2): 226–232.
Zou, Y., Tian, M., and Liu, C. (2016). Optimization of ultrasound-assisted extraction of puerarin from Pueraria
lobata dried root. J Food Process Preserv. 40: 431–436.
ChaptEr 10
Use of pulsed Light in Food processing
Lakshmi E. Unni and O. P. Chauhan
CONtENtS
10.1 Introduction ......................................................................................................................... 173

10.2 Principle .............................................................................................................................. 174
10.3 Generation of Pulsed Light ................................................................................................. 175
10.4 Mechanism of Action .......................................................................................................... 177
10.5 Factors Affecting the Microbial Inactivation by Pulsed Light ........................................... 178
10.5.1 Type of Microorganism ......................................................................................... 178
10.5.2 Food Composition ................................................................................................. 178
10.5.3 Distance from the Light Source ............................................................................ 180
10.5.4 Applications in Food Industry............................................................................... 180
10.5.4.1 Fruits and Vegetables ........................................................................... 180
10.5.4.2 Dairy Products...................................................................................... 182
10.5.4.3 Animal Products and Sea Food ............................................................ 183
10.5.5 Effect on Functional and Nutritional Properties ................................................... 183
10.5.6 Merits and Demerits of Pulsed-Light Technology ................................................ 184
10.5.6.1 Merits.................................................................................................... 184
10.5.6.2 Demerits ............................................................................................... 184
10.6 Conclusion and Future Strategies ....................................................................................... 184
References ...................................................................................................................................... 185
10.1 INtrODUCtION
Food safety is one of the most critical issues currently affecting the public health system.
Over the last few decades, technology and innovation have contributed to an apparent reduction
in the mortality rates resulting from exposure to improperly processed and/or handled food. These
improvements correspond to industry’s implementation of preventive controls (Hazard Analysis and
Critical Control Points) in the manufacturing, distribution, and holding of foods and food ingre-
dients. Changes in consumption and taste across global markets also pose challenges in the food
manufacturing sector. The conventional approach of a food processor is to develop products with
a long shelf-life in user-friendly packaging systems. They are usually bothered about sensory attri-
butes and microbiological profiles without much emphasis on the fate of heat, labile, nutritional, and
nutraceutical components. As consumers are looking for more and more nutritive foods, preservation
technologies that are more efficient from the microbiological point of view, but that are not capable
173
of causing losses in the sensory and nutritional characteristics of foods, are becoming well known in
this context (Cruz et al., 2011). A series of advanced physical technologies have been proposed over
the past years but their further up-scaling and industrial application require a better understanding of
their mechanisms of antimicrobial action and efficacy in controlling food safety. A few years before,
non-thermal techniques were restricted to framing of fundamentals with regard to the inactivation
of vegetative and reproductive bodies of microbes and the deteriorative enzymes. Ever since then,
food engineering has taken rapid strides towards the design and development of process machinery
to achieve commercialization of these techniques. High-intensity pulsed light (PL) has demonstrated
potential for use in food safety and food hygiene control applications. Over the past 20 years, a
number of researchers have investigated pulsed light as a means for decontaminating foods and food
contact surfaces, non-opaque liquids, and packaging materials.
PL is a technique to decontaminate surfaces by killing microorganisms using short time pulses of
an intense broad spectrum, rich in UV-C light. UV-C is the portion of the electromagnetic spectrum
corresponding to the band between 200 and 280 nm. PL employs light wavelengths ranging from
ultraviolet (UV) to near infrared in short intense pulses (Palmieri and Cacace, 2005). The intensity of
PL is at least 20,000 times more than that of sun light (Bushnell et al., 1998). Pulses of light used for
food processing applications typically emit one to twenty flashes of electromagnetic energy per sec-
ond. The material to be treated is exposed to at least one pulse of light having an energy density in the
range of about 0.01 to 50 J/cm2 at the surface. A wavelength distribution such that at least 70% of the
electromagnetic energy is within the range from 170 to 2600 nm is used. PL refers to broad-spectrum
radiation delivered as intense, intermittent, short-duration pulses. A typical PL lamp produces poly-
chromatic radiation in the wavelength range of 100–1100 nm, although the wavelength range can vary
based on the lamp source. It is done with the help of Xenon lamps that can produce flashes several
seconds (Green et al., 2005). PL is also referred to as high-intensity light, broad-spectrum white light,
intense PL, pulsed white light, and pulsed UV light. PL is effective for sterilization of surfaces and
highly transparent liquids, such as water, and has been successfully used to inactivate bacteria, mould,
spores, and viruses in various food materials. This technology had been successfully used to inactivate
myriad microorganisms in various foods. UV-C treatment for preserving food was discovered in the
1930s (Artes and Allende, 2005). The use of PL for the inactivation of microorganisms was initi-
ated in Japan in the late 1970s. Later extensive work was done to sterilize pharmaceutical products.
However, the technology was adopted by the food industry in 1996, only when the Food and Drug
Administration Authority approved the use of PL technology for production, processing, and handling
of foods. PL is considered a non-thermal technology, even though there is a significant increase in
temperature, because this increase is a by-product of treatment. PL is also an effective alternative to
the chemical sterilants frequently used for treating food packaging materials, as it does not produce
chemical residues. For example, consider a food product that is incompatible with hydrogen peroxide
(H2O2) because of its tendency to develop rancidity as a result of contact with residues remaining on
packaging materials treated with H2O2. In this context, PL serves as a viable method for decontamina-
tion of packaging (Ohlsson and Bengtsson, 2002).
10.2 prINCIpLE
The electromagnetic radiations, emitted and propagated as waves, are characterized by their
wavelength (λ), frequency (f), and energy (E). The term light refers to radiations having λ ranging
about from 180 to 1100 nm, which includes UV rays (λ = 180–400 nm, roughly subdivided into
UVA, 315–400 nm, UVB, 280–315 nm, UVC, 180–280 nm), visible light (λ = 400–700 nm), and
infrared rays (IR, λ = 700–1100 nm) (Figure 10.1). Light can be emitted from different sources by
different mechanisms, due to the spontaneous transition of some atoms from an excited state to a
condition of lower energy. The excited atoms can either (i) relax back to ground state by releasing
use oF PuLseD LiGHt in FooD ProCessinG 175
Figure 10.1 Pulsed-light spectrum.
energy as heat, (ii) relax back to the ground state by releasing energy as photons, or (iii) induce some
chemical changes. The microorganisms undergo these changes on UV exposure and get inactivated
due to chemical changes and temperature build-up (Demirci and Krishnamurthy, 2011). Light can be
delivered either continuously or in the form of pulses (Palmieri and Cacace, 2005). The most impor-
tant feature of delivering energy in the form of pulses is that apart from the number and duration of
pulses, power provided by the pulses is greater than that provided by a continuous light radiation of
equivalent total energy; total energy being equal, the shorter the duration of each pulse, the higher
the pulse power. For this reason, if compared with continuous light radiations, light pulses show a
much higher penetrating capability through the materials (Dunn et al., 1989). Within a fraction of
a second, the electromagnetic energy gets stored in the capacitor and is then released in the form of
light within a billionth of a second, which results in power amplification and minimum additional
energy consumption (Green et al., 2005). The inactivation efficacy of PL depends upon intensity
(measured in J/cm2) and the number of pulses delivered. This technology is always characterized by
the use of following units:
Fluence rate: It is the energy received from the lamp by the sample per unit area per second. Its unit
is Watt/meter2 (W/m2).
Fluence/dose: It is the energy received from the lamp by the sample per unit area during the treatment.
Its unit is Joule/meter2 (J/m2).
Pulse width: It is the time interval (fractions of seconds) during which energy is delivered.
Exposure time: It is the time period in seconds during which treatment is given.
Peak power: It is measured as pulse energy divided by the pulse duration. Its unit is Watt (W).
Pulse-repetition-rate (prr): It is the number of pulses per second (Hertz [Hz]) or commonly expressed
as pps (pulses per second).
10.3 GENEratION OF pULSED LIGht
PL system consists of several common components (Figure 10.2). Equipment used to produce PL
is made up of a power unit, one or more adjustable xenon lamp units, and a high-voltage connection
that allows the transfer of a high-current electrical pulse (Figure 10.3). A high-voltage power supply
provides electrical power to the storage capacitor, which stores electrical energy for the flash lamp.
The flash lamps are usually filled with xenon or krypton inert gases due to their high efficiency of
electrical to optical energy conversion and emit broadband radiation that ranges from UV to near
infrared. As the current passes through the gas chamber of the lamp unit, the electrons surrounding
the Xe atoms are excited, causing them to jump to higher energy levels. The electrons release this
energy and drop back to a lower energy level by emitting photons. The wavelength distribution of
the optical spectrum emitted by xenon flash lamp ranges from 100 to 1,100 nm: UV (100–400 nm),
visible light (400–700 nm), and infrared (700–1,100 nm). Many fluids, such as water, have a high
degree of transparency to a broad range of wavelengths including visible and UV light, while other
AC power Switch
input
Pulsed Light source

Capacitor
Rectifier DC Power
Food
Treatment chamber
Figure 10.2 schematic diagram of pulsed-light generation.
2 5
7
1
0.5
1. Power and Control module
2. Control cable
3. UV sterilization chamber
4. Cooling duct
5. Blower
6. UV lamps
1 3
7. Sample holder
Figure 10.3 Pulsed-light system.
liquids, such as sugar solutions and wines, exhibit a more limited transparency. Increasing the amount
of solids will diminish the intensity of penetration of the UV radiation. The lethal effect of PL can
be attributed to the rich broad-spectrum UV content, its short duration, high peak power, and the
ability to regulate both the pulse duration and the frequency output of the flash lamp. In the case of
aseptic packaging system, the packaging may be spread with light-enhancing agents and subjected
to the light pulses. The containers are passed through several stations while being disinfected. The
disinfected containers are filled with processed commercially sterile food products which are sub-
sequently sealed at the top with a sterile lid. The lids may also be disinfected using light pulses. The
food product flows through the chamber surrounding the PL source. The treatment chamber may
be designed to include a reflector assembly as an outer wall or external reflection to reflect back the
illumination traversing the food product (Dunn et al., 1995).
10.4 MEChaNISM OF aCtION
As a substantial portion of the PL spectrum covers UV light, the antimicrobial effects of PL are
well known to be primarily mediated by its UV light component. The latter is absorbed by highly
conjugated double-bond systems in biomolecules, leading to their chemical modification. It was
also found that there is no killing effect if a filter is used to remove UV wavelength region lower
than 320 nm (Takeshita et al., 2003). The UV spectrum comprises three wave ranges: long-wave
UV-A (320–400 nm), medium-wave UV-B (280–320 nm), and short-wave UV-C (200–280 nm).
Mechanisms that have been proposed to explain the lethality of PL treatment are related to the
shortwave UV-C part of the spectrum (Anderson et al., 2000; Takeshita et al., 2003; Wuytack
et al., 2003). The effects of pulsed UV light on microbial cells can be classified as photochemical
(e.g., thymine-thymine dimer formation in microbial DNA), photo-thermal (localized heating
of bacteria), and photo-physical (constant disturbance caused by the high-energy pulses) effects.
However, their relative importance depends on the fluence and target microorganism. The effects
of UV light on microbes are attributed to structural changes in the DNA, as well as abnormal
ion flow, increased cell membrane permeability, and depolarization of the cell membrane (Gomez
Lopez et al., 2007). UV light absorbed by the conjugated carbon-carbon double bonds in proteins
and nucleic acids induces the antimicrobial effect as it changes the DNA and RNA structures.
Due to the absorption of the UV light, cross-linking between neighboring pyrimidine nucleoside
bases (thymine and cytosine) in the same DNA strand occurs. Formation of the hydrogen bonds
to the purine bases on the opposite strand is impaired due to the mutated base. DNA transcription
and replication is thereby blocked, compromising cellular functions and eventually leading to cell
death. The amount of cross-linking is proportional to the amount of UV exposure. The levels of
mutations that can be reversed depend on the UV repair system present in the target microorganism.
Once the threshold of cross-linking has been exceeded, the number of cross-links is beyond repair,
and cell death occurs (Miller et al., 1999). In the UV-C range of 250–260 nm, alterations in DNA
take place due to pyrimidine dimers mainly thymine dimers (Mitchell et al., 1992; Giese and Darby,
2000). UV irradiation usually generates thymine dimers in large quantity, cytosine dimers in low
quantity, and mixed dimers at an intermediate level (Figure 10.4) (Setlow and Carrier, 1966). These
dimers inhibit the formation of new DNA chains in the process of cell replication resulting in the
chologenic death of affected microorganisms by UV (Bolton and Linden, 2003). It was also found
by experiments that enzymatic repair of DNA does not occur after damaged by PL. Enzymes are
among the major targets for photo-induced modifications due to the abundance of endogenous chro-
mophores within their structure. Both amino acid side-chains (for example, tryptophan, tyrosine,
phenylalanine, cysteine) and bound prosthetic groups (e.g., flavins, heme) may act as efficient chro-
mophores. Enzymatic proteins have the additional ability to bind exogenous chromophores, and
rapidly react with other excited state species (Davies and Truscott, 2001). The result would thus be
a modification in the protein properties due to side-chain oxidation, backbone fragmentation, and/
or formation of cross-links and aggregates.
The lethal effect of PL can also be due to photo-thermal effect. Most energy of the light pulses
that penetrate through a food product is absorbed by the layers nearest to the surface and dissipated
as heat, causing in such thin layers a certain increase in temperature. Since microbial cells have a
higher absorption of the PL than that of the surrounding medium (water), this determines a localized
rapid heating of microorganisms, which at very high pulse power values can reach temperatures
(about 130°C) sufficient to cause their overheating, rupture, and death and the extremely short pulse
duration prevents the microorganism cell surface from being cooled by the surrounding medium
(Wekhof, 2000). It was proposed that with a fluence exceeding 0.5 J/cm2, the disinfection is achieved
through a rupture of bacteria during their temporary overheating caused by absorption of all UV
light from a flash lamp. The ruptured top of the spore shows evidence of an escape of an overheated
Figure 10.4 Formation of thymine dimer, thymine-cytosine dimer and mixed dimer. (Adapted from setlow, r.
and Carrier, W.L., J. Mol. Biol., 17, 237–254, 1966.)
content of the spore, which became empty after such an internal “explosion” and “evacuation” of
its content took place during the light pulse. Other effects on the cells include collapse of cell struc-
ture and enlargement of vacuoles as found in some microbial studies (Proctor, 2011) as showed by
flashed yeast cells. As PL causes cell membrane damage, it could be considered as a technique for
sterilization (Takeshita et al., 2003). The effectiveness of PL treatment depends upon several factors
such as intensity, treatment time, food temperature, and type of microorganism. Table 10.1 repre-
sents a brief summary of microbial inactivation studies conducted in several foods.
The susceptibility trend is reported to be gram-negative bacteria > gram-positive bacteria >
bacterial spores > fungal spores (Rowan et al., 1999; Anderson et al., 2000; Levy et al., 2012).
10.5 FaCtOrS aFFECtING thE MICrOBIaL INaCtIVatION BY pULSED LIGht
10.5.1 type of Microorganism
Optical properties of cells, for example, their degree of scattering and absorption of light, are
important. The incident beam of light undergoes refraction due to difference in the optical density
between the substrate and the surrounding air. There are also some micro-organisms resistant to PL
(Ethan, 2009; Rajkovic et al., 2010).
10.5.2 Food Composition
Food composition also affects the efficacy of the decontamination by PL. Gómez-Lopez et al.
(2005b) treated Photobacterium phosphoreum, L. monocytogenes, and Candida lambica inocu-
lated onto surfaces of agars supplemented with several food components. The results demonstrated
that proteins and oil decreased the decontaminant efficacy of PL, whereas when water or starch
was added to the agar, no particular trends were observed. Roberts and Hope (2003) also found
that the addition of protein to a buffered saline solution decreased the efficacy of virus inactivation.
Therefore, high protein and fat-containing food products have little potential to be efficiently treated
table 10.1 Microbial Inactivation by pulsed Light in Foods

Experimental
Microorganism Substrate Condition Log reduction references
pathogenic Bacteria
Bacillus cereus Agar F = 3; t = 20 4.9 rowan et al. (1999)
F = 7; t = 1500 3.0 Gómez-Lopez et al. (2005a)
Bacillus circulans Agar F = 7; n = 1500 3.7 Gómez-López et al. (2005a)
Clostridium Agar F = 7; t = 1500 >2.9 Gómez-López et al. (2005a)
perfringens
Salmonella enteritidis Agar F = 3; t = 20; 5.6 rowan et al. (1999)
n = 200
Klebsiella oxytoca Agar F = 7; t = 50 4.2 Gómez-López et al. (2005a)
Shigella flexnii Agar F = 7; t = 1500 3.8 Gómez-López et al. (2005a)
Salmonella Agar F = 7; t = 1500; 3.2
typhimurium n = 50
Escherichia coli Agar F = 3; t = 20 6.2 rowan et al. (1999)
O157:H7
F = 7; t = 1500 4.7 Gómez-López et al. (2005a)
F = 3; t = 512 6.82 MacGregor et al. (1998)
Yersina enterocolitica Agar F = 7; t = 1500 3.9 Gómez-López et al. (2005a)
Staphyloccocus Agar F = 5.6; 8.5 Krishnamurthy et al. (2004)
aureus buffer solution t = 5400 7.5
Agar
Spoilage Bacteria
Alicyclobacillus Agar F = 7; t = 1500 >5.2 Gómez-López et al. (2005a)
acidoterrestris
Bacillus circulans Agar F = 7; t = 1500 >4.1 Gómez-López et al. (2005a)
Lactobacillus sake Agar F = 7; t = 1500 2.5 Gómez-López et al. (2005a)
Photobacterium Agar F = 7; t = 1500 >4.4 Gómez-López et al. (2005a)
phosphoreum
Pseudomonas Agar F = 3; t = 20; 6.8 rowan et al. (1999)
aeruginosa n = 200
Pseudomonas Agar F = 7; t = 1500; 4.2 Gómez-López et al. (2005a)
fluorescens n = 50
Fungi
Aspergillus flavus Agar F = 7; t = 1500 2.2 Gómez-López et al.
(2005a)
Aspergillus niger buffer solution F = 1; t = 1000; 4.8 Wekhof et al. (2001)
n=5
Botrytis cinerea Agar F = 7; t = 1500 1.2 Gómez-López et al. (2005a)
Fusarium culmorum Agar F = 3; t = 85 4.5 Anderson et al. (2000)
Monilia fructigena Agar F = 7; t = 1500 4 Marquenie et al. (2003)
Yeasts
Saccharomyces Agar F = 3; t = 20; 4.9 rowan et al. (1999)
cerevisiae n = 100
buffer solution F = 0.7; 6.0 takeshita et al. (2003)
t = 1200; n = 5
Candida lambica Agar F = 7; t = 1500; 2.8 Gómez-López et al. (2005a)
n = 50
Rhodotorula Agar F = 7; t = 1500; >2.8 Gómez-López et al.
mucilaginosa n = 50 (2005a)
Note: F is the Fluence (J/cm2); t is the treatment time (μs); n is the number of pulses.
by PL. Vegetables, on the other hand, could therefore be suitable for PL treatment. For transparent
and colored food materials, refraction is particularly relevant, whereas for opaque food materials,
reflection is the prevailing phenomenon. Specular or diffused reflection can occur depending on
the smoothness or roughness of the surface of the material respectively. For smooth surfaces, the
incident light bounces on the surface and comes out at the same angle as the incident beam. For
rough surfaces, light travels through the outer layers of the material, where the incident light is
partly absorbed. For translucent materials, some part of the incident light interacts with the internal
structures and causes multiple internal reflections and redirections, which result in scattering. In the
case of biological tissues, absorption and scattering are the most relevant types of light–substrate
interaction (Cheong et al., 1990).
10.5.3 Distance from the Light Source
As the distance from light source and depth of the substrate increases, the absorption and
scattering diminishes. This is because the light intensity decreases as it travels through the sub-
strate. The quantitatively distribution of light dose inside a substrate is described by the term optical
penetration depth, which represents the distance over which light decreases in fluence rate to 37% of
its initial value. The optical penetration varies with wavelength, with shorter wavelengths providing
deeper penetration into the food than longer wavelengths (Dagerskog and Osterstrom, 1979).
10.5.4 applications in Food Industry
In the food industry, PL can be applied to sterilize, sanitize, or reduce microbial load in foods and food
packaging materials, as well as surfaces, environments, plants, devices, and media (water, air) involved
in food processes. The effectiveness of PL is strongly influenced by the interaction of the substrate with
the incident light. Therefore, the treatment is most effective on smooth, non-reflecting surfaces or in
clear liquids. Overall, PL treatment brings new opportunities to the food industry, ranging from water
disinfection to the manufacture of safe, non-heat-treated fruit juices, surface treatment of foods, and food
contact materials, or the terminal antimicrobial treatment of foods packaged in UV transparent materi-
als. This technology could be used as a means to increase the safety and shelf life of foods, with no det-
rimental effects on their quality and sensory properties. PL has potential applications in food processing
that requires a rapid disinfection where surface contamination is a concern for microbial contamination
such as fresh whole fruit and vegetable commodities, hard cheeses, meat slices, and so on (Bhavya and
Umesh, 2017). Table 10.2 represents the effect of PL processing on various foods.
10.5.4.1 Fruits and Vegetables
Dunn et al. (1989) treated potato slices by 2–5 flashes of broad-spectrum PL at a fluence of
30 kJ/m2 and observed that they retained their color after a prolonged storage, while the untreated
samples began to brown through the action of polyphenol oxidase in a few minutes; however, the
opposite unexposed surfaces of treated samples showed visible browning. Similar effects have been
demonstrated when treating slices of bananas and apples. Manzocco et al. (2011) demonstrated
UV-C light exposure may allow for the non-thermal decontamination of the surface of fresh-cut
melon, thus extending its shelf-life. Ignat et al. (2014) demonstrated that PL may allow non-thermal
decontamination of the surface of fresh-cut apple slices with negligible changes in fresh-like
appearance of the product. Aguilo-Aguayo et al. (2013) reported that PL was able to reduce by
about 1 log natural and inoculated microbial contamination on tomato fruit and did not lead to loss
of nutritional quality, as vitamin C was unaffected while carotenoid concentrations were actually
table 10.2 Effect of pulsed-Light processing on Foods
product Experimental Condition Microorganism Log reduction references

Liquid Foods
Apple juice Frequency 3 pulses/s and E. coli AtCC 25922 2.66 log CFu/mL sauer and
pulse width 360 μs: fluence Moraru (2009)
12.6 J/cm2 E. coli o157: H7 2.52 log CFu/mL
Apple cider E. coli AtCC 25922 2.32 log CFu/mL
E. coli o157: H7 3.22 log CFu/mL
orange juice 200–1100 nm, 3 Hz and E. coli 2.90 log CFu/mL Palgan et al.
36 μs, 1.17 J/cm2/pulse at a L. innocua 0.93 log CFu/mL (2011)
distance of 2.5 cm:
7–28 J/cm2
Milk 200–1100 nm, 3 Hz and E. coli 0.61–1.06 log CFu/mL Palgan et al.
36 μs, 1.17 J/cm2/pulse at a L. innocua 0.51–0.84 log CFu/mL (2011)
distance of 2.5 cm:
7–28 J/cm2
F = 25 J/cm2; n = 110 Serratia >2.0 log CFu/mL smith et al.
marcescens (2002)
3 pulses/s and 1.27 J/cm2/ Staphylococcus 0.55–7.23 log CFu/mL Krishnamurthy
pulse aureus et al. (2007)
Others Foods
Corn meal F = 5.6 J/cm2; n = 300 Aspergillus niger 4.93 log CFu/mL Jun et al.
spores (2003)
Wheat flour F = 1.95 J/cm2; n = 64 Saccharomyces 0.7 log CFu/mL Fine and
cerevisiae Gervais
(2004)
Alfalfa seeds F = 5.6 J/cm2; n = 135 Escherichia coli 0.94–1.82 log CFu/mL sharma and
Demirci
(2003)
black pepper F = 1.95 J/cm2; n = 64 Saccharomyces 2.93 log CFu/mL Fine and
cerevisiae Gervais
(2004)
Honey F = 5.6 J/cm2; n = 135 Clostridium 0.89–5.46 log CFu/mL Hillegas and
sporogenes Demirci
(2003)
Sea Foods
salmon fillets F = 5.6 J/cm2; n = 135 Listeria 0.72–0.8 log CFu/mL ozer and
monocytogenes Demirci
(2005)
salmon fillets F = 5.6 J/cm2; n = 135 Escherichia coli 0.24–0.91 log CFu/mL ozer and
Demirci
(2005)
shrimp fillets F = 6.3–12.1 J/cm2 L. monocytogenes 2.2–2.4 log CFu/g Cheigh et al.
Flatfish fillets 1.9–2.1 log CFu/g (2013)
salmon fillets 1.7–1.9 log CFu/g
Fruits and Vegetables
strawberries F = 7 J/cm2; n = 3750 Botrytis cinera 3.0 log CFu/mL Marquenie
et al. (2003)
F = 7; n = 3750; Monilia fructigena 4.0 log CFu/mL
t = 30 μs
F = 64.8 J/cm2 ; n = 180 Escherichia coli 3.3 log CFu/mL bialka and
O157:H7 Demirci
(2008)
(Continued)
table 10.2 (Continued) Effect of pulsed-Light processing on Foods
product Experimental Condition Microorganism Log reduction references

raspberries F = 59.4; n = 180 Salmonella spp. 3.4 log CFu/mL bialka and
Demirci
(2008)
blueberries F = 5–5.61 J/cm2; E. coli O157:H7 3.8–>6.7 log CFu/g Huang and
t = 360 μs Salmonella 4.8–5.7 log CFu/g Chen (2014)
Apple F = 17.5–157.5 kJ/m2; Listeria 2.72–3.86 log CFu/cm2 ignat et al.
t = 50 μs monocytogenes (2014)
Lactobacillus brevis 2.15–2.90 log CFu/cm2
Plum F = 5.4 J/cm2 B. cereus 1.4 log CFu/g Luksiene et al.
(2012)
tomato t = 112 μs 1.5 log CFu/g
Cauliflower 1.3 log CFu/g
spinach F = 8 J/cm2 L. innocua 1.85 log CFu/g Aguero et al.
(2016)
E. coli 1.72 log CFu/g
slightly increased. Charles et al. (2013) investigated the impact of PL treatment on physical and
nutritional quality of fresh-cut mangoes and found by flashing samples at 8 J/cm2 that color, firm-
ness, but also nutritional qualities are maintained. PL showed a marked influence in reducing the
microbial spoilage of PL-treated fresh cut avocados and could extend their shelf-life up to 15 days
(Aguilo-Aguayo et al., 2014). Pineapple sticks submitted to 200 J/m2 UV-C light, packaged in con-
ventional PET/EVOH/PE trays and stored at 6°C up to 15 days showed slower yeast and lactic acid
bacteria growth (Manzocco et al., 2016). Sauer and Moraru (2009) achieved a 7.15-log cfu reduction
of Escherichia coli in apple juice. Raspberries treated with PL showed a decrease in brightness and
did not substantially change the redness of the fruit immediately after the treatment, but along the
storage, berries became darker red and decrease in brightness, and a similar trend in the stability of
firmness of the fruit was also observed (Xu and Wu, 2016).
Gómez-Lopez et al. (2005a) achieved a range of 0.5–2.04-log reduction of mesophilic, aerobic
microbes naturally found in minimally processed vegetables such as spinach, iceberg lettuce, cab-
bage, celeriac, green bell peppers, and soybean sprouts, using a treatment dose of up to 2700 light
pulses. An 80% increase in the respiration rate of shredded iceberg lettuce with no significant effects
on the respiration rate of shredded white cabbage treated under the same conditions was observed.
Koh et al. (2016) noticed that a decrease in the pH and an increase in acidity were more pronounced
for untreated samples compared to PL-treated fresh-cut cantaloupes, throughout the storage.
10.5.4.2 Dairy Products
A high fluence of 26.25 J/cm2 resulted in 3.2 log reduction in the total microbial count, and
concomitantly, milk temperature was increased to 55°C, which indicated a combined effect of pho-
tochemical and photo-thermal damage of natural microflora by PL in raw milk (Innocente et al.,
2014). In cheddar cheese, PL did not affect the color and lipid peroxidation during refrigerated
condition, but panellists scored the PL-treated samples lower than the untreated ones for the sen-
sory attributes such as overall liking, flavor, and appearance. However, a dose of 9.22 J/cm2 had
an adverse effect on organoleptic properties of cheese (Proulx et al., 2017). Similarly, a significant
difference in odor and flavor in the cheese slices treated with 4.2 and 8.4 J/cm2 shows the presence
of sulphur notes and the difference in decontamination magnitude between types of cheese, which
could be explained by their different topography that is porous nature in the Manchego- vs. smooth
in the Gouda-types of cheeses (Fernandez et al., 2016).
10.5.4.3 Animal Products and Sea Food
Hierro et al. (2012) conducted the sensory analysis and found that PL fluences of 8.4 and
4.2 J/cm2 or lower did not affect the raw attributes such as odor and color of beef and tuna carpaccio,
respectively. However, during shelf-life studies, beef and tuna carpaccio showed significant, remark-
able difference in color and odor when treated with PL at 4.2 J/cm2 and above. Changes in a* and
b* values were observed for RTE loin samples compared to Salchichon along the 28 days storage
when PL of 11.9 J/cm2 was applied. Color parameters were not dramatically modified by PL treat-
ment in these RTE dry-cured products, which may be attributed to the greater stability of the cured
pigments in comparison to those of fresh meat (Ganan et al., 2013). High-power PL of 1,000 pulses,
treatment duration 200 seconds, and total UV light dose 5.4 Joule/cm2 was found to reduce viability
of Salmonella typhimurium and Listeria monocytogenes inoculated on the surface of chicken by
2–2.4 log10 (N/N0) colony-forming units (CFU)/mL (Paskeviciute et al., 2011).
Eggs were treated with PL of flashes of 2.1 and 10.5 J/cm2. Exposure to 2.1 J/cm2 leads to death
of Salmonella cells (5 log colony-forming units (CFU) per egg shell) on the egg surface with slight
increase in temperature. Increase to 10.5 J/cm2 did not cause penetration of Salmonella cells to the
egg contents from the shell. No adverse effect on quality of egg albumin was observed. No effect on
sensory and functional properties (Lasagabster et al., 2011).
Ozer and Demirci (2005) have reported a 1.09-log reduction for Listeria monocytogenes on
raw salmon fillets after 180 pulses of light. Figueroa-Garcia et al. (2002) found that fresh cat-
fish fillets treated by 2–4 pulses of 2.5–5 kJ/m 2 showed a slightly increased oxidative rancidity
(thiobarbituric acid oxidation values) after 2 days of refrigerated storage, but such values did not
differ from those of untreated samples thereafter and did not reach typical levels of rancidity for
sea foods.
10.5.5 Effect on Functional and Nutritional properties
Since the wavelengths used for PL are too long to cause ionisation of small molecules and
are in the non-ionising portion of the electromagnetic spectrum (Dunn et al., 1995), the forma-
tion of radioactive by-products is not expected. Light-induced flavor is caused due to activation
of riboflavin which is responsible for the conversion of methionine from methanol which leads to
burnt protein-like or medicinal-like flavor. Peroxides produced using UV light exposure may also
attack fat-soluble vitamins and colored compounds and may lead to change in nutritional quality
and discoloration. Furthermore, long treatments with UV light may increase the temperature of
food product, which will lead to temperature-related quality changes such as cooked flavor and
change in color due to non-enzymatic browning. UV light can also degrade vitamins (especially
A, C, B2) by photo-degradation. UV radiation may also denature proteins, enzymes, and amino
acids in milk, leading to textural changes. Water also absorbs UV photons and produces OH− and
H+ radicals which in turn aids change in other food components. Though UV light exposure when
applied at high doses affects the structural stability of food components, most of the changes are
detrimental to microbial growth. Normally, microbial inactivation can be achieved within seconds
to minutes depending upon the opacity of food products and microorganism type. Therefore, food
quality may not be affected negatively when the process is done at optimized conditions (Demirci
and Krishnamurthy, 2011). Apples treated with PL were susceptible to surface browning when com-
pared to untreated samples (Moreira et al., 2017). PL induces degradation of biopolymers in cell
wall, affects the pectin present in the cell wall, and cells appeared collapsed with ruptured mem-
branes and thus causing a rupture and folding of cell walls. This membrane damage would increase
in enzymatic browning reactions like polyphenol oxidase activity due to greater tissue damage and
loss of functional cell compartmentalization (Gomez et al., 2012b). Gomez et al. (2012a) also found
changes in total profile analysis, dynamic, and creep behavior on apple surface due to PL treatment
and storage period. The use of PL at high fluences (28 J/cm2) dramatically affects the final quality
of fresh-cut mushrooms, and thermal damage due to high PL doses seems to cause dehydration and
major textural modification (Oms-Oliu et al., 2010).
10.5.6 Merits and Demerits of pulsed-Light technology
10.5.6.1 Merits
The inactivation of microbes by PL is a very fast process and causes a rapid disinfection in
a very short period. It is a green technology as the consumption of energy is very less during its
application. It has been proven as a safe technology for living beings and their environment without
producing harmful residuals, chemicals and toxic by-products in the PL-treated foods. The con-
cerns of ionized radicals and radioactive by-products in foods by consumers are removed in PL due
to its non-ionizing spectrum. The intensity of light, which lasts for only a second, is 20,000 times
brighter than sunlight, but there is no thermal effect, so quality and nutrient content are much
retained (Brown, 2008). The xenon-flash lamps used in PL treatment are eco-friendlier than the
mercury vapor lamps used in UV treatment (Gomez-Lopez et al., 2007). Compared with some
alternative techniques that use chemical preservatives or ionizing radiations, PL technology does
not involve toxic chemicals or photolytic by-products (the used wavelengths are too long to cause
ionization of small molecules). Furthermore, if compared with conventional continuous UV light
treatment (UVCL), PL, because of its higher power and short duration, allows more effective micro-
bial inactivation to be achieved, requiring a relatively lower energy input, penetrating opaque or
thick materials better than UVCL, and causing significantly lower product thermal damage. It is
claimed that PL systems have relatively low operating costs and do not significantly contribute to
the environmental impact because they do not produce volatile organic compounds or suspended
airborne particulates, and they generate little liquid and solid wastes.
10.5.6.2 Demerits
PL application in meat industry has some constraints as it has low penetration power and chances
of lipid oxidation. To get the desired outcome, the packaging materials showing high penetration of
PL should be used while treating the packaged food in this method. The limited control of food heat-
ing still remains the main concern in PL technology. Sample heating is perhaps the most important
limiting factor of PL for practical applications. Another drawback of this method is that folds or fis-
sures in the food may protect microbes from being exposed to the PL (Brown, 2008). There might be
some strains of micro-organisms which are resistant to the PL treatment, for example Listeria mono-
cytogenes (Caminiti et al., 2011). This technique for decontamination of micro-organisms is useful
mostly in case of liquid foods and surface of solid foods, hence limiting its application. Another
disadvantage of PL treatment is the possibility of shadowing occurring when microorganisms readily
absorb the rays, as in the case of A. niger, and are present one upon another. This makes the organ-
isms in the lower layers very hard to destroy in contrast to those in the upper layer (Hiramoto, 1984),
although the use of relatively high peak powers can overcome the shadowing effect.
10.6 CONCLUSION aND FUtUrE StratEGIES
PL technology holds considerable promise in food processing to be used for pasteurization of

juices and beverages, as a post lethality treatment in controlling microbial contamination on meats
and shell eggs surfaces, and as a means for the shelf-life extension of fresh produce. In order to mini-
mize the thermal effect, slight alteration is required by the addition of cooling systems. However, to
improve the efficacy of UV light for food application, the following areas of research need to be con-
ducted. A more comprehensive understanding is need of inactivation mechanisms of gram-positive
and gram-negative bacteria by PL, how the fluence, type of microbial strain, as well as physical and
chemical properties of the foods affects the occurrence of lethal and sublethal effects induced by
PL treatment. Furthermore, kinetic inactivation data need to be obtained for pathogen and spoilage
microorganisms, taking into account interactions between microorganisms and surface materials,
such as shielding effects from incident UV and their dependency on surface structure or topography.
Thus, more sound studies in the indicated areas can ensure the effectiveness of UV light for micro-
bial inactivation, stimulate the growing interest in the non-thermal technologies, and could bridge
gap between research achievements and commercialization of this technology.
rEFErENCES
Aguero, M. V., Jagus, R. J., Martin-Belloso, O., and Soliva-Fortuny, R. (2016). Surface decontamination of spinach
by intense pulsed light treatments: Impact on quality attributes. Postharvest Biol. Technol. 121: 118–125.
Anderson, J. G., Rowan, N. J., MacGregor, S. J., Fouracre, R. A., and Farish, O. (2000). Inactivation of food-
borne enteropathogenic bacteria and spoilage fungi using pulsed-light. IEEE Trans. Plasma Sci. 28(1):
83–88.
Artes, F., and Allende, A. (2005). Processing lines and alternative preservation techniques to prolong the shelf
life of minimally processed leafy vegetables. Eur. J. Hortic. Sci. 70: 231–245.
Aguilo-Aguayo, I., Charles, F., Renard, C. M., Page, D., and Carlin, F. (2013). Pulsed light effects on sur-
face decontamination, physical qualities and nutritional composition of tomato fruit. Postharvest Biol.
Technol. 86: 29–36.
Aguilo-Aguayo, I., Oms-Oliu, G., Martín-Belloso, O., and Soliva-Fortuny, R. (2014). Impact of pulsed light
treatments on quality characteristics and oxidative stability of fresh-cut avocado. LWT-Food Sci.
Technol. 59(1): 320–326.
Bhavya, M. L., and Umesh Hebbar, H. (2017). Pulsed light processing of foods for microbial safety. Food
Qual. Saf. 1(3): 187–202.
Bialka, K. I., and Demirci, A. (2008). Efficacy of pulsed UV-light for the decontamination of Escherichia coli
O157:H7 and Salmonella enterica on raspberries and strawberries. J. Food Sci. 73(5): M201–M207.
Bolton, J. R., and Linden, K. G. (2003). Standardization of methods for fluence (UV dose) determination in
bench-scale UV experiments. J. Environ. Eng. 129: 209–215.
Brown, A. C. (2008). Understanding Food: Principles and Preparation, p. 47. Thompson/Wadsworth
Publishing, Belmont, CA.
Bushnell, A., Cooper, J. R., Dunn, J., Leo, F., and May, R. (1998). Pulsed light sterilization tunnels and sterile-
pass-throughs. Pharmaceutical Eng. 18(2): 48–58.
Caminiti, I. M., Noci, F., Munoz, A., Whyte, P., Morgan, D. J., and Cronin, D. A. (2011). Impact of selected
combinations of non-thermal processing technologies on the quality of an apple and cranberry juice
blend. J. Food Chem. 124(4): 1387–1389.
Charles, F., Vidal, V., Olive, F., Filgueiras, H., and Sallanon, H. (2013). Pulsed light treatment as new method
to maintain physical and nutritional quality of fresh-cut mangoes. Innov. Food Sci. Emerg. Technol. 18:
190–195.
Cruz, R. M. S., Rubilar, J. F., Ulloa, P. A., Torres, J. A., and Vieira, M. C. (2011). New food processing technol-
ogies: Development and impact on the consumer acceptability. pp. 555–584. In: Food Quality: Control,
Analysis and Consumer Concerns, Medina, D. A., and Laine, A. M., Eds., Nova Science Publishers,
Hauppauge, NY.
Cheigh, C. I., Hwang, H. J., and Chung, M. S. (2013). Intense pulsed light (IPL) and UV-C treatments for inac-
tivating Listeria monocytogenes on solid medium and seafoods. Food Res. Int. 54: 745–752.
Cheong, W., Prahl, S. A., and Welch, A. J. (1990). A review of the optical properties of biological tissues. IEEE
J. Quantum Electron. 26(12): 2166–2185.
Dagerskog, M., and Osterstrom, L. (1979). Infra-red radiation for food processing I: A study of the fundamen-
tal properties of infra-red radiation. Lebensm. Wissen. Technol. 12: 237–242.
Davies, M. J., and Truscott, R. J. (2001). Photo-oxidation of proteins and its role in cataractogenesis.
J. Photochem. Photobiol. 63(1–3): 114–125.
Demirci, A., and Krishnamurthy, K. (2011). Pulsed ultraviolet light. In: Nonthermal Processing Technologies
for Food, pp. 249–261. Zhang, H. Q., Barbosa-Cánovas, G. V., Balasubramaniam, V. M., Dunne, C. P.,
Farkas, D. F., and Yuan, J. T. C., Eds., Wiley-Blackwell, Hoboken, NJ.
Dunn, J., Ott, T., and Clark, W. (1995). Pulsed light treatment of food and packaging. Food Technol. 49(9):
95–98.
Dunn, J. E., Clark, W. R., and Asmus, J. F. (1989) Methods for preservation of foodstuffs. Maxwell Laboratories,
San Diego, CA. US Patent 4871559.
Ethan, S. (2009). The Produce Contamination Problem: Causes and Solutions. p. 280. Gerald, M. S., and
Matthews, K. R., Eds., Academic Press, San Diego, CA.
Fernandez, M., Arias, K., and Hierro, E. (2016). Application of pulsed light to sliced cheese: Effect on Listeria.
Food Bioprocess Technol. 9: 1335–1344.
Fine, F., and Gervais, P. (2004). Efficiency of pulsed UV light for microbial decontamination of food powders.
J. Food Prot. 67: 787–792.
Figueroa-Garcia, J. E., Silva, J. L., Kim, T., Boeger, J., and Cover, R. (2002). Use of pulsed-light to treat raw
channel catfish fillets. J. Miss. Acad. Sci. 47(2): 114–120.
Ganan, M., Hierro, E., Hospital, X. F., Barroso, E., and Fernandez, M. (2013). Use of pulsed light to increase
the safety of ready-to-eat cured meat products. Food Control 32: 512–517.
Giese, N., and Darby, J. (2000). Sensitivity of microorganisms to different wavelengths of UV light:
Implications on modeling of medium pressure UV systems. Water Res. 34(16): 4007–4013.
Gomez, P. L., Garcia-Loredo, A., Nieto, A., Salvatori, D. M., Guerrero, S., and Alzamora, S. M. (2012a).
Effect of pulsed light combined with an antibrowning pretreatment on quality of fresh cut apple. Innov.
Food Sci. Emerg. Technol. 16: 102–112.
Gomez, P. L., Salvatori, D. M., Garcia-Loredo, A., and Alzamora, S. M. (2012b). Pulsed light treatment of
cut apple: Dose effect on color, structure, and microbiological stability. Food Bioprocess Technol. 5:
2311–2322.
Gómez-Lopez, V. M., Devlieghere, F., Bonduelle, V., and Debevere, J. (2005a). Intense light pulses decon-
tamination of minimally processed vegetables and their shelf-life. Int. J. Food Microbiol. 103: 79–89.
Gómez‐Lopez, V. M., Devlieghere, F., Bonduelle, V., and Debevere, J. (2005b). Factors affecting the inactiva-
tion of micro‐organisms by intense light pulses. J. Appl. Microbiol. 99(3): 460–470.
Gomez-Lopez, V. M., Ragaert, P., Debevere, J., and Devlieghere, F. (2007). Pulsed light for food decontamina-
tion: A review. Trends Food Sci. Technol. 18(9): 464–473.
Green, S., Basaran, N., and Swanson, B. (2005). Food Preservation Techniques. p. 365. In: Zenthen, P., and
Bogh-Sorenson, L., Eds., Woodhead Publishing House, CRC Press, Washington, DC.
Hierro, E., Ganan, M., Barroso, E., and Fernandez, M. (2012). Pulsed light treatment for the inactivation of
selected pathogens and the shelf-life extension of beef and tuna carpaccio. Int. J. Food Microbiol. 158:
42–48.
Hillegas, S. L., and Demirci, A. (2003). Inactivation of Clostridium sporogenes in clover honey by pulsed
UV-light treatment. In 2003 ASAE Annual Meeting, p. 1. American Society of Agricultural and
Biological Engineers, Saint Joseph, MI.
Hiramoto, T. (1984). Method of sterilization. Ushio Denki Kabushikikaisha, Tokyo, Japan. US Patent 4464336.
Huang, Y., and Chen, H. (2014). A novel water-assisted pulsed light processing for decontamination of blue-
berries. Food Microbiol. 40: 1–8.
Ignat, A., Manzocco, L., Maifreni, M., Bartolomeoli, I., and Nicoli, M. C. (2014). Surface decontamination
of fresh-cut apple by pulsed light: Effects on structure, colour and sensory properties. Postharvest Biol.
Technol. 91: 122–127.
Innocente, N., Segat, A., Manzocco, L., Marino, M., Maifreni, M., Bortolomeoli, I., and Nicoli, M. C.
(2014). Effect of pulsed light on total microbial count and alkaline phosphatase activity of raw milk.
Int. Dairy J. 39: 108–112.
Jun, S., Irudayaraj, J., Demirci, A., and Geiser, D. (2003). Pulsed UV-light treatment of corn meal for inactiva-
tion of Aspergillus niger spores. Int. J. Food Sci. Technol. 38: 883–888.
Krishnamurthy, K., Demirci, A. L. I., and Irudayaraj, J. (2004). Inactivation of Staphylococcus aureus by
pulsed UV-light sterilization. J. Food Prot. 67(5): 1027–1030.
Krishnamurthy, K., Demirci, A., and Irudayaraj, J. M. (2007). Inactivation of Staphylococcus aureus in milk
using flow-through pulsed UV-light treatment system. J. Food Sci. 72(7): M233–M239.
Koh, P. C., Noranizan, M. A., Karim, R., and Nur Hanani, Z. A. (2016). Microbiological stability and quality
of pulsed light treated cantaloupe (Cucumis melo L. reticulatus cv. Glamour) based on cut type and light
fluence. J. Food Sci. Technol. 53: 1798–1810.
Lasagabster, A., Arboleya, J. C., and Martinez, D. M. (2011). Pulsed light technology for surface decontami-
nation of eggs: Impact on Salmonella inactivation and egg quality. Innov. Food Sci. Emerg. Technol.
12(2): 124–128.
Levy, C., Aubert, X., Lacour, B., and Carlin, F. (2012). Relevant factors affecting microbial surface decon-
tamination by pulsed light. Int. J. Food Microbiol. 152: 168–174.
Luksiene, Z., Buchovec, I., Kairyte, K., Paskeviciute, E., and Viskelis, P. (2012). High-power pulsed light for
microbial decontamination of some fruits and vegetables with different surfaces. J. Food Agri. Environ.
10: 162–167.
MacGregor S. J., Rowan N. J., Mcllvaney L., Anderson J. G., Fouracre R. A., and Farish O . (1998). Light inactiva-
tion of food-related pathogenic bacteria using a pulsed power source. Letters in App. Microbiol. 27: 67–70.
Manzocco, L., Da Pieve, S., and Maifreni, M. (2011). Impact of UV-C light on safety and quality of fresh-cut
melon. Innov. Food Sci. Emerg. Technol. 12: 13–17.
Manzocco, L., Plazzotta, S., Maifreni, M., Calligaris, S., Anese, M., and Nicoli, M. C. (2016). Impact of UV-C
light on storage quality of fresh-cut pineapple in two different packages. LWT-Food Sci. Technol. 65:
1138–1143.
Marquenie, D., Michiels, C. W., Van Impe, J. F., Schrevens, E., and Nicolai, B. N. (2003). Pulsed white light
in combinations with UV-C and heat to reduce storage rot of strawberry. Postharvest Biol. Technol. 28:
455–461.
Miller, R. V., Jeffrey, W., Mitchell, D., and Elasri, M. (1999). Bacterial responses to ultraviolet light. ASM
News 65: 535–541.
Mitchell, D. L., Jen, J., and Cleaver, J. E. (1992). Sequence specificity of cyclobutane pyrimidine dimers in
DNA treated with solar (ultraviolet B) radiation. Nucleic Acids Res. 20(2): 225–229.
Moreira, M. R., Alvarez, M. V., Martin-Belloso, O., and Soliva-Fortuny, R. (2017). Effects of pulsed light
treatments and pectin edible coatings on the quality of fresh-cut apples: A hurdle technology approach.
J. Sci. Food Agri. 97: 261–268.
Ohlsson, T., and Bengtsson, N. (2002). Minimal processing of foods with non-thermal methods. In: Minimal
Processing Technologies in the Food Industry, p. 112. Ohlsson, T., and Bengtsson, N., Eds., Woodhead
Publishing, CRC Press, Cambridge, UK.
Oms-Oliu, G., Aguilo-Aguayo, I., Martin-Belloso, O., and Soliva-Fortuny, R. (2010). Effects of pulsed
light treatments on quality and antioxidant properties of fresh-cut mushrooms (Agaricus bisporus).
Postharvest Biol. Technol. 56: 216–222.
Ozer, N. P., and Demirci, A. (2005). Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes
inoculated on raw salmon fillets by pulsed UV‐light treatment. Int. J. Food Sci. Technol. 41(4): 354–360.
Palgan, I., Caminiti, I. M., Munoz, A., Noci, F., Whyte, P., Morgan, D. J., and Lyng, J. G. (2011). Effectiveness
of high intensity light pulses (HILP) treatments for the control of Escherichia coli and Listeria innocua
in apple juice, orange juice and milk. Food Microbiol. 28: 14–20.
Palmieri, L., and Cacace, D. (2005). High intensity pulsed light technology. In: Emerging Technologies for
Food Processing, pp. 279–306. Sun, D., Ed., Academic Press, New York.
Paskeviciute, E., Buchovec, I., and Luksiene, Z. (2011). High Power Pulsed light for decontamination of
chicken from food pathogens: A study on antimicrobial efficiency and organoleptic properties. J. Food
Saf. 31: 61–68.
Proctor, A. (2011). High intensity pulsed light food processing. In: Alternatives to Conventional Food
Processing, p. 200. Proctor, A., Ed., Royal Society of Chemistry (RSC), Fayetteville, AR.
Proulx, J., Agustin, M., Sullivan, G., VanWees, S., Jian, J., Hilton, S. T., and Moraru, C. I. (2017). Short com-
munication: Influence of pulsed light treatment on the quality and sensory characteristics of cheddar
cheese. J. Dairy Sci. 100: 1004–1008.
Rajkovic, A., Tomasevic, I., Smigic, N., Uyttendaele, M., Radovanovic, R., and Devlieghere, F. (2010). Pulsed
ultraviolet light as an intervention strategy against Listeria monocytogenes and Escherichia coli
O157:H7 on the surface of a meat slicing knife. J. Food Eng. 100: 446–451.
Roberts, P., and Hope, A. (2003). Virus inactivation by high intensity broad spectrum pulsed light. J. Virol.
Methods. 110: 61–65.
Rowan, N. J., MacGregor, S. J., Anderson, J. G., Fouracre, R. A., McIlvaney, L., and Farish, O. (1999). Pulsed-
light inactivation of food-related microorganisms. J. Appl. Environ. Microbiol. 65: 1312–1315.
Sauer, A., and Moraru, C. I. (2009). Inactivation of Escherichia coli ATCC 25922 and Escherichia coli O157:
H7 in apple juice and apple cider, using pulsed light treatment. J. Food Prot. 72(5): 937–944.
Setlow, R., and Carrier, W. L. (1966). Pyrimidine dimers in ultraviolet-irradiated DNA’s. J. Mol. Biol. 17(1):
237–254.
Sharma, R. R., and Demirci, A. (2003). Inactivation of Escherichia coli O157:H7 on inoculated alfalfa seeds
with pulsed ultraviolet light and response surface modelling. J. Food Sci. 68: 1448–1453.
Smith, W. L., Lagunas-Solar, M. C., and Cullor, J. S. (2002). Use of pulsed ultraviolet laser light for the cold
pasteurization of bovine milk. J. Food Prot. 65(9): 1480–1482.
Takeshita, K., Shibato, J., Sameshima, T., Fukunaga, S., Isobe, S., and Arihara, K. (2003). Damage of yeasts
induced by pulsed light irradiation. Int. J. Food Microbiol. 85: 151–158.
Wekhof, A. (2000). Disinfection with flash lamps. PDA J. Pharm. Sci. Technol. 54(3): 264–276.
Wuytack, E. Y., Phuong, L. D. T., Aertsen, A., Reyns, K. M. F., Marquenie, D., De Ketelaere, B., and Michiels,
C. W. (2003). Comparison of sublethal injury induced in Salmonella enterica serovar Typhimurium by
heat and by different nonthermal treatments. J. Food Prot. 66(1): 31–37.
Xu, W., and Wu, C. (2016). The impact of pulsed light on decontamination, quality, and bacterial attachment
of fresh raspberries. Food Microbiol. 57: 135–143.
ChaptEr 11
Ozone application in Food processing
C. T. Manoj Kumar and Latha Sabikhi
CONtENtS
11.1 Introduction ......................................................................................................................... 190

11.2 Synthesis and Molecular Structure of Ozone ..................................................................... 191
11.3 Physico-chemical Characteristics of Ozone........................................................................ 191
11.3.1 Solubility ................................................................................................................ 191
11.3.2 Stability .................................................................................................................. 192
11.4 Generation of Ozone ........................................................................................................... 193
11.4.1 Electrical Discharge Method (Corona Discharge) ................................................. 193
11.4.2 Electrochemical Method (Cold Plasma Method) ................................................... 193
11.4.3 Radiation Method................................................................................................... 193
11.4.4 Mechanism of Microbial Inactivation .................................................................... 194
11.5 Factors Affecting Activity of Ozone ................................................................................... 194
11.5.1 Temperature ........................................................................................................... 194
11.5.2 pH ........................................................................................................................... 195
11.5.3 Organic Components.............................................................................................. 196
11.5.4 Type of Microorganisms and Their Initial Load ................................................... 196
11.6 Application of Ozone in Selected Food Commodities........................................................ 196
11.6.1 Fruits and Vegetables ............................................................................................. 196
11.6.1.1 Effect on Microbial Safety .................................................................... 196
11.6.1.2 Effect on Quality ................................................................................... 197
11.6.1.3 Effect on Nutrients ................................................................................ 197
11.6.1.4 Effect on Pesticide Residue ................................................................... 198
11.6.2 Fruit Juices ............................................................................................................. 199
11.6.3 Grain Storage .........................................................................................................200
11.6.3.1 Effect on Pests or Insects.......................................................................200
11.6.3.2 Effect on Microorganisms ..................................................................... 201
11.6.3.3 Effect on Mycotoxins ............................................................................ 201
11.6.3.4 Effect on Quality ...................................................................................202
11.6.4 Meat........................................................................................................................202
11.6.4.1 Effect on Microorganisms .....................................................................202
189
11.6.5 Fish .........................................................................................................................204

11.6.5.1 Effect on Microorganisms .....................................................................204
11.7 Safety and Risks ..................................................................................................................206
References ......................................................................................................................................206
11.1 INtrODUCtION
Food safety is the prime factor for any food processing industry to have potential impact on
public health, food security, and trade competitiveness. Outbreaks of food spoilage lead to serious
health problems and end up with great economic losses. Conventional thermal food processing
techniques involved in assuring food safety induce adverse effects on the natural characteristics
and nutritional value of food. Hence, the food processing industry is under continuous pressure
to produce fresh, minimally processed, natural, healthy, and safe foods. Non-thermal processing
of food material is a potential objective to meet the above requirements. Various non-thermal
processes can be classified into physical, chemical, and biological methods. The commonly used
physical methods such as high-pressure processing, ultrasound, and others are still in their infancy
and they are limited in the food processing industry due to their high capital and maintenance
costs. The chemicals used in disinfection/detoxification of food materials leave toxic residues
when they come in contact with organic food constituents. Commonly used chemicals such as
chlorine, fluorine, and iodine (halogens) produce trihalomethane, a carcinogenic compound.
Also, ammonia, sodium bisulfite, calcium hydroxide, formaldehyde, and hydrogen peroxide, used
against fungal contaminants, also release toxic residues. Ozonation is a disinfection process where
the triatomic oxygen (ozone, O3) has been used as active agent in decreasing microbial content.
The antimicrobial effect of ozone is due to its strong oxidizing effect (2.07 mV); it leaves no resi-
dues after processing and mainly produces no toxic compounds during processing. Hence, it has
been recommended in various disinfection applications. It achieved “generally regarded as safe”
(GRAS) status decades back from the United States Food and Drug Administration (USFDA) for
use as disinfectant in drinking water and during wastewater treatments. Later, in 1997, an expert
panel convened by the Electric Power Research Institute approved the GRAS status for ozone
for use as a direct disinfectant or sanitizer in foods. After some brief investigation, finally FDA
granted GRAS status to ozone (USFDA, 2001). Ozone has been used in numerous food products
as an effective disinfectant. The specific advantage of ozone over the common disinfectants is
that it leaves no toxic residues after processing. The application of ozone in water treatment is
widespread throughout the world. Now, it has been successfully commercialized in household
drinking water purifiers and is also recommended for waste water treatments for sedimentation of
suspended materials, chemical, and biological oxidation. Among food materials, it was success-
fully applied on fruit and vegetables, food grains, meat, and meat products as well as fish products
and its derivatives to enhance their shelf stability without affecting their quality and nutritional
characteristics (Karaca and Velioglu, 2007; Tiwari et al., 2010; Pohlman, 2012; Goncalves, 2016).
Ozone has been used as an antifungal agent in cereal grains and also for destruction of mycotox-
ins such as aflatoxins (Tiwari et al., 2010). The oxidation property of ozone was also utilized to
improve the functional properties of proteins and starch by modifying their structure (Uzun et al.,
2012). However, its application in dairy industry has been restricted to cleaning of processing lines
and removing residues and biofilm-forming bacteria on surface of stainless steel. Strong oxidative
property of ozone causes oxidation of milk fat causing unfavorable oxidizing flavor production
(Varga and Szigeti, 2016). As such, ozonation is a cost effective and environmentally friendly
food-processing technology.
oZone APPLiCAtion in FooD ProCessinG 191
11.2 SYNthESIS aND MOLECULar StrUCtUrE OF OZONE
Ozone is a triatomic oxygen molecule and its molecular structure renders strong oxidizing powers
to ozone molecule. During synthesis of ozone molecule, the oxygen molecule will be broken into two
unpaired oxygen atoms (called as homolytic fission, free radical), each one occupying one 2p orbital.
O2 UV light or High electric arc

 → 2O ( with two unpaired electron )
This unpaired electron containing oxygen atom (O−, free radical) combines with one oxygen
molecule (O2) to form ozone (O3).
O2 + O − → O3 ( Unstable)
During ozone formation three oxygen atoms are combined, where exactly, in central oxygen a 2s
and two 2p (2pz and 2py) atomic orbitals are rearranged to form one 2sp2 molecular orbital (hybrid
orbital). This 2sp2 orbital of one oxygen (O1) atom combined with 2sp2 of central oxygen atom (O2)
and one 2sp2 orbital of another oxygen atom (O3) combined with 2p2 orbital of central oxygen atom
(O2) leads to formation of triatomic oxygen molecule (ozone). These arrangements exhibit triangle
shape with an angle of 116° 49′ and both the atoms are at the same distance with bond length of
1.278 Å. As a consequence, the ozone molecule represents a hybrid formed by the four canoni-
cal structures (Figure 11.1). The high reactivity of ozone can then be attributed to the absence of
electrons in one of the terminal oxygen atoms in some of the symmetric structures confirming the
electrophilic character of ozone, while the excess negative charge present in some other oxygen
atom imparts a nucleophilic character.
11.3 phYSICO-ChEMICaL CharaCtErIStICS OF OZONE
11.3.1 Solubility
Solubility property of ozone is dependent on several extrinsic factors such as temperature, pres-
sure, composition of the medium, pH, gas flow rate, etc. Henry’s law says that the pressure of the gas
exerts above the liquid is directly proportional to the solubility of the ozone (Bablon et al., 1991). De
Smedt et al. (2001) revealed that solubility of ozone is dependent on pH, temperature, and type of
additives of the aqueous solution (Table 11.1). Levanov et al. (2008) found that solubility of ozone
decreased with the addition of phosphoric acid and perchloric acid at 20°C. In case of sulfuric acid,
the solubility is minimum at 12 M concentration and then increases; at 17.9 M, it reaches equal to
water. The solubility ratio (ratio between the concentrations of ozone in solution and gas phase) was
0.276 in pure water, 0.122 in 12 M, and 0.265 in 17.9 M sulfuric acid. Horvath et al. (1985) found
temperature-dependent solubility ratio data varied from 0.31 to 1.13. Huge variation in the solubility
ratio was also found in several research efforts, which may be attributed to the different analytical
methods, reactor design, and gas flow rate (Kim et al., 2003). Numerous studies have been carried
out to measure the solubility of ozone and expressed in Henry’s law constant (K H), pseudo Henry’s
Figure 11.1 structure of the ozone molecule as a resonance hybrid of the four canonical forms.
table 11.1 physico-chemical properties of Ozone

Characteristics
Colour blue (when generated from ordinary air)
Colourless (from pure air)
Dark blue liquid at −112°C
odour Pungent
Melting point −192.5 ± 0.4°C
boiling point −111.9 ± 0.3°C
Critical temperature −12.1°C
Critical pressure 54.6 atm
Density 2.14 g/L at 0°C and atm pressure
oxidation-reduction potential 2.07 V
Molecular weight g/mole 47.9982
solubility in water, ppm at 20°C 3
Heat of formation kJ/mole 144.7
specific gravity 1.658
table 11.2 temperature and Solubility relationship of Ozone in Water

temperature (°C) Solubility (Litre Ozone/Litre Water)
0 0.640
15 0.456
27 0.270
40 0.112
60 0.000
law constant, and Ostwald coefficient (Battino et al., 1983). Generally, ozone is partially soluble in
water. However, temperature plays a major role in solubility of ozone, as temperature increases solu-
bility decreases. At 0°C, ozone solubility is 0.640 L ozone/L water, whereas at 60°C it is insoluble
in water (Hill and Rice, 1982). Its solubility in water is 13 times that of oxygen at 0°C–30°C and
it is progressively more soluble in colder water (Rice, 1986). Also, according to the Henry-Dalton
constants, the solubility of the ozone is higher at lower temperature (Table 11.2).
11.3.2 Stability
Ozone is a most unstable molecule, easily dissociating into oxygen molecule and hydroxide ion.
Ozone readily degrades but has a longer half-life in the gaseous state than in aqueous solution. Gao
et al. (2008) reported that purity of water, pH, and temperature usually affects ozone stability. As
the conductance and half-life of the water increased the ozone stability will also increase, whereas
increase in temperature and pH decreased the stability of ozone. Impurities present in the solutions
even worsen the degradation of ozone more than pure water. Pan et al. (1984) studied the effect of
pH, temperature, buffer solutions, and minerals on ozone decomposition. The study reported that
ozone is reasonably stable at moderately low pH (~3 pH) and ambient temperatures in acetic, sulfu-
ric, and nitric acid solutions. Its decomposition was slightly higher at 15°C and moderately higher at
35°C than 25°C in all three buffer solutions. Effect of minerals was found to depend on concentra-
tion, particularly at 0.5 ppm concentration only, whereas Co (II) ion enhanced the decomposition
of ozone in sulfuric acid solution. However, at the higher concentration level (∼3.0 ppm), Ca(II),
Cr(III), Fe(II), Fe(III), Co(II), Ni(II), and Cu(II) ions were found to contribute to the decomposition
of ozone; the effect of Co(II) and Fe(II) ions was very pronounced as compared to the other ions.
Sulfuric acid as buffer reagent showed drastic and moderate reductions of ozone decomposition
compared to acetic acid solutions, which is attributed to the radical scavenging mechanism of acetic
acid. Yang and Chen (1979) reported that the stability of ozone in water depends on water tempera-
ture, initial ozone concentration, and length of holding time. In general, ozone was more stable in
water at 2°C than in water at 25°C. Percentages of ozone retained after standing were similar for
initial concentrations, which ranged from 3.15 to 4.65 mg/L.
11.4 GENEratION OF OZONE
Ozone can be generated by exposing pure oxygen gas to a highly electrical energy source or
ultraviolet radiation, where oxygen molecule (O2) splits into individual oxygen atom (O−) and com-
bines with another oxygen molecule (O2) to form ozone (O3). Since it is an unstable gas that quickly
decomposes, the half-life of the ozone is about 20–30 min at 20°C (Khadre et al., 2001), and hence,
it should be generated onsite for immediate use. Naturally, ozone is formed in the stratosphere by
high-ultraviolet radiations sparks, but for commercial use it can be generated by employing electrical
discharge (high-voltage electrical arc), electrochemical, and radiation methods. Among these, electri-
cal discharge is the most widely used commercial method; however, it is less efficient (2%–10%) and
demands more electrical energy over electrochemical and radiation methods (Mahapatra et al., 2005).
11.4.1 Electrical Discharge Method (Corona Discharge)
The electrical discharge method comprises two parallel electrodes separated by dielectric mate-
rials, where high voltage has been employed between them. The air or pure oxygen gas used as
input material, it should be dried to a dew point of at least −60°C and free from particulate matters.
When high voltage is passed in electrodes, a corona discharge is developed between two electrodes;
the oxygen gas present in this gap get split off into oxygen atoms and is then combined with other
oxygen molecule to form ozone and get discharged as ozone/mixture of gas. A corona discharge is
a physical phenomenon characterized by a low-current electrical discharge across a gas-containing
gap at a voltage gradient, which exceeds a certain critical value (Taylor et al., 1996). During ozone
generation, the temperature of the gap will be increased, which is associated with degradation
of generated ozone. In order to prevent the generation of heat during ozone formation, coolant
(air or water) has to be circulated. The discharged gas mixture normally contains 1%–3% of ozone
when dry air is used and 3%–6% of ozone when pure oxygen gas is used (Mahapatra et al., 2005).
Rice et al. (1981) claimed that 16% of ozone can be generated using pure oxygen gas as input.
11.4.2 Electrochemical Method (Cold plasma Method)
In electrochemical method of ozone production, anode and cathode placed in an electronega-

tive anions solution, an electric current is passed between them to generate ozone at the anode. The
method is advantageous over other methods as it consumes low-voltage DC current, no feed gas is
required, it requires less surface area for installation, and it has an expected high yield of ozone
(Mahapatra et al., 2005).
11.4.3 radiation Method
In radiation method, both ultraviolet and gamma radiations are used for production of ozone.
In ultraviolet method of O3 generation, the ozone is formed when O2 is exposed to UV light of
140–190 nm wavelength, which splits the oxygen molecules into oxygen atoms; then the oxygen
atoms combine with other oxygen molecules to form O3 (Muthukumarappan et al., 2000). However,
due to poor yields, this method has very limited uses.
11.4.4 Mechanism of Microbial Inactivation
Extensive research work has been done on evaluation of antimicrobial activity of ozone and it
has been well documented in several review papers and book chapters. Ozone has a strong anti-
microbial nature on a wide range of microorganisms including gram-positive and gram-negative
microorganisms, spores, and vegetative cells (Guzel-Seydim et al., 2004; Greene et al., 2012).
The high oxidizing power of ozone and its decomposed products (free radicals) are responsible
for the strong antimicrobial nature, where it acts on glycolipids, glycoproteins of cell wall, cell
membrane, enzyme system, and nucleic acids of the microorganisms. Two main mechanisms are
widely quoted in literature: the oxidizing power of ozone affects the sulfhydryl groups and amino
acids of enzymes (Al-Haddad et al., 2005; Torlak et al., 2013), as well as peptides, proteins, and
polyunsaturated fatty acids of cell membrane (Victorin, 1992). As the surface affected leads to
formation of pores in the cell wall and cell membrane associated with expulsion of cell material
to the outside, and also bulging of the cell due to movement of water from outside to inside, it
leads to cell bursting and causes cell death. Microbial cell death is also associated with destruction
of nucleic acids by ozone (Kim et al., 1999). Destruction of DNA caused by change in its second-
ary structure (Hunt and Marinas, 1999) or mutation (Torlak et al., 2013) leads to change in genetic
material. Thymine is more sensitive than cytosine and uracil. Ozone also destroys viral RNA and
alters polypeptide chains in viral protein coats (Kim et al., 1999). The destruction of microbial struc-
ture upon ozone treatment has been described by Alwi and Ali (2014) and is given in Figure 11.2.
11.5 FaCtOrS aFFECtING aCtIVItY OF OZONE
11.5.1 temperature
Temperature is the major factor in affecting the antimicrobial efficacy of ozone. Several
researchers worked on effects of temperature on ozone efficacy and found contradictory and
inconclusive results. Torlak (2014) found that the antimicrobial effect of ozone on Alicyclicbacillus
acidoterrestris spores in apple juice at 4°C was higher as compared to 22°C. It may be attributed
to increased solubility and decomposition rate of ozone, as the temperature increases decom-
position rate increases and there will be no free ozone to act on microbials. Hence, temperature
should be kept low during application of ozone, so that availability of ozone is increased and,
consequently, its efficacy (Rice et al., 1981; Pascual et al., 2007). Similar results have been
obtained when ozone is used for inactivation of E. coli O157:H7 and Salmonella in apple cider
and orange juice (Williams et al., 2004). Even the decomposed ozone releases free radicals
but they are less effective in microbial inactivation than molecular ozone (Zuma et al., 2009).
On the other hand, some researchers found apparently contradictory results. Steenstrup and
Floros (2004) reported that in ozonated apple cider D values of E. coli O157:H7 was increased
with decreasing temperature. As temperature increases the decomposition rate of ozone also
increases, the released free radicals exert higher oxidization power than ozone molecule, and,
therefore, as the temperature increases the inactivation also increases (Manousaridis et al.,
2005). The varied antimicrobial activity of ozone due to the temperature would be attributed to
the kind of experimental setup followed by the researchers (Kim et al., 2003). Also, it is depen-
dent on type of microorganism likely to react with either ozone molecule or free radicals to get
inactivated (Blatchley and Hunt, 2002).
Figure 11.2 scanning electron micrographs of control (a, c, and e) and ozonated (b, d, and f) cultures of E. coli
o157, Salmonella typhimurium and L. monocytogenes, respectively. note that the cells of control
bacterial cultures have smooth shape and intact (iC) while the cells of treated bacterial cultures
has irregular shape and disrupted (DC). (Adapted from Alwi, n.A. and Ali, A., Food Control, 46,
304–311, 2014.)
11.5.2 ph
The pH is one of the intrinsic factors of the food medium which affects the antimicrobial activity
of ozone. The acidic pH favours the faster inactivation of microorganisms by ozone compared to
alkaline pH (Patil et al., 2010; Song et al., 2015). Hence, the acidic nature of fruit juices improves
antibacterial effectiveness of ozone and it has been considered as the suitable media for ozonation
(Torlak, 2014). Reduction in antimicrobial effectiveness at higher pH may be attributed to faster
decomposition of ozone into free radicals (Patil et al., 2010) and also as such reduced pH aid in
inhibition of growth of microorganisms.
11.5.3 Organic Components
The organic components are the constituents of the food products such as proteins, minerals,
and antioxidants which are suspected to be involved in affecting the antimicrobial activity of ozone
(Choi et al., 2012). Restaino et al. (1995) reported that the addition of bovine serum albumin reduces
the residual ozone content as well as antimicrobial activity of ozone, whereas addition of soluble
starch did not show any effect. Choi et al. (2012) reported that increase in solids content of juice
significantly reduces the ozone antimicrobial activity. It may be attributed to chelation of ozone by
antioxidant and minerals present in fruit juice.
11.5.4 type of Microorganisms and their Initial Load
The sensitivity of microorganisms to ozone is dependent on their structural characteristics. For

example, bacteria are more sensitive to ozone than yeasts and fungi. While gram-positive bacteria
are more sensitive than gram-negative bacteria (Greene et al., 2012). It may be due to low degree of
crosslinking of cell wall in gram-positive bacteria (Navarre and Schneewind, 1999), which reduced
its rigidity and increased susceptibility to ozone (Alwi and Ali, 2014). However, Tzortzakis and
Chrysargyris (2017) reported that gram-positive bacteria are more resistant to ozone than gram-negative
bacteria, because gram-positive bacteria have more thicker cell membrane and hydrophilic than gram-
negative bacteria. In case of spores and vegetative cells, spores are more resistant to ozone because of
their thicker multilayer protein protection (Greene et al., 2012; Tzortzakis and Chrysargyris, 2017).
Additionally, the ozone antimicrobial activity is dependent on initial microbial load, as microbial
population increases the antimicrobial activity of ozone decreases. It may be attributed to decomposi-
tion of ozone into oxygen molecule (O2) enhancing the growth of residual microorganisms.
11.6 appLICatION OF OZONE IN SELECtED FOOD COMMODItIES
11.6.1 Fruits and Vegetables
Fruits and vegetables are the most perishable commodity having demand throughout the year. To
meet the demand generated on a daily basis there is a need to preserve the fruits and vegetables from
spoilage and pathogenic organisms as long as possible. Ozonation as a surfaced infection process
can be applied effectively to these perishable commodities immediately after harvesting without
affecting the nutritive value and other quality characteristics. Numerous reports are available on eval-
uation of antimicrobial nature of ozone both in gaseous and aqueous form on fruits and vegetables.
11.6.1.1 Effect on Microbial Safety
Fruits are easily deteriorated with bacterial and fungal contamination and ozone has been used
as surface disinfectant in many fruits either in gaseous or in aqueous form.
Horvitz and Cantalejo (2012) studied that application of gaseous ozone at the concentration of
0.7 μmol mol−1 was a more efficient sanitizer than aqueous ozone. The antimicrobial efficacy of
ozone against pathogenic bacteria such as E. coli, Listeria monocytogenes, and Salmonella have
been determined on several fruits such as fresh-cut bell peppers (Alwi and Ali, 2014) and fresh-cut
papaya (Yeoh et al., 2014). Alwi and Ali (2014) reported that Listeria monocytogenes was most
sensitive to ozone treatment, and Yeoh et al. (2014) reported that coliforms were more sensitive
than mesophiles. However, reduction in count was very less indicating the antimicrobial activity of
ozone. A minimum 2 log CFU g−1 reduction of microbial count can be considered as antimicrobial
agent (Torlak et al., 2013). However, Ketteringham et al. (2006) reported that the fresh-cut peppers
washed with ozonated water did not observe any reduction in microbial counts. They suggested that
treating whole peppers is better than precut pepper, because leaching of organic matter from precut
peppers may interfere with ozone action. Alwi and Ali (2014) claimed it as resistance offered by
bacterial cells against ozone activity.
The antifungal activity of ozone in fruit processing has been assessed on citrus fruits (orange
and lemon) (Palou et al., 2001), where ozone treatment did not stop the growth of blue or green
mould (Penicillium digitatum and Penicillium italicum) but delayed it by 1 week. Yaseen et al.
(2014) observed that the raspberries, strawberries, and blueberries treated with high concentration
of ozone (2000 nL.L−1) prior to storage showed antifungal activity similar to that of continuous
exposure during storage at low concentration of ozone (300 nL.L−1). Whangchai et al. (2006) stud-
ied the post-harvest decay of the longan fruit treated with gaseous ozone of 200 μl.L−1 for 0, 15,
30, 60, and 120 min and stored at 25°C. They reported that the fruit treated for 120 min reduced the
surface microflora population noticeably; however, the disease incidence was increased in the fruit
exposed to 120 min as compared to 60 min. They suggested that the longer exposure to ozone dam-
ages the cellular membrane. Fungal hyphae penetrate into the fruit through lenticels, making fruit
more susceptible to decay. Application of ozone gas in controlling the sporulation of Penicillium
digitatum and P. italicum in oranges packed in citrus cartons, naked, or bagged returnable plastic
containers (RCPs) or bagged in fiber board master cartons (Palou et al., 2003). The sporulation was
significantly inhibited by ozone exposure only on oranges packed naked in RPCs compare to other
packed samples because of high penetration of ozone in RPCs.
11.6.1.2 Effect on Quality
Quality characteristics of fruits and vegetables are the decisive factor during their marketing. The
loss of water, firmness, and colour as the consequence of senescence of reduce their marketing value
and they become more susceptible to mechanical damage during handling. Hence, several research-
ers worked on the effect of ozone in protecting the quality characteristics during storage. Glowacz
et al. (2015) studied the effect of ozone (0.1 and 0.3 μmol mol−1) on quality changes during storage
of red bell peppers, cucumbers, and zucchini stored at 14°C, 12°C, and 8°C, respectively. Weight
loss, color, and texture of the peppers showed no significant changes during storage at both ozone
concentrations. In cucumbers and zucchini, continuous exposure to 0.1 μmol mol−1 showed weight
loss reduction by more than 40% and improvement in textural maintenance. Minas et al. (2012) stud-
ied the effect of gaseous ozone (0.3 μl.L−1) on physicochemical properties (firmness, soluble solids,
phenolics, and ascorbic acid content) of kiwi fruit to address the post-harvest ripening of kiwi fruit.
They observed that firmness was retained even after 5 months of storage and ethylene production was
slowed down even after 5 months of storage; the authors believed that the ozone might have blocked
the climacteric ethylene rise in the fruit reducing the softening of fruit. The ascorbic acid and phenol
content were increased after one day of storage. Similar results obtained in case of papayas treated
with ozone (0, 1.5, 2.5, 3.5, and 5 ppm for 96 h). The firmness of the papayas retained during 10 days
of storage at 25°C. The percent of water loss was increased with increase in the days of storage,
whereas increase in the concentration of ozone did not show any effect. The lightness of the fruit
increases during fruit ripening and decreases once the fruit becomes over ripened. The ozone treat-
ment was found to reduce the papaya fruit ripening by 2 days (Ali et al., 2014).
11.6.1.3 Effect on Nutrients
Fruits are rich source of nutrients such as minerals, dietary fibers, vitamins, and antioxidants.
Application of ozone on whole or fresh-cut fruit pieces has been reported to affect the nutrients
adversely. The literature available on effect of ozone treatment on nutritional value of the fruit and
vegetables majorly targeted the antioxidants and vitamins, whereas, the effect on other nutrients
such as dietary fibers, minerals, and carbohydrates are not much emphasized. The fruit and veg-
etables are rich source of vitamin C or ascorbic acid. The ozone treatment showed mixed trend
on vitamin C content of fruits and vegetables. The vitamin C was reported to decrease in apples
(Swami et al., 2016), pineapple, banana, guava (Alothman et al., 2010), and kiwi fruit (Minas et al.,
2012). Reduction in ascorbic acid content by exposing to ozone for long time may be due to the
radical scavenging mechanisms initiated by ascorbic acid against free products (free radicals) of
auto-decomposed ozone. It was found that activation of ascorbate oxidase may cause degradation
of ascorbic acid. Ascorbate oxidase is copper-containing enzyme responsible for enzymatic deg-
radation of ascorbic acid into dehydroascorbic acid (DHA). Similarly, cyanidine 3-glucoside in
apple decreased after treatment with ozone (Swami et al., 2016). However, some researchers found
increase in vitamin C content in red bell peppers (Glowacz et al., 2015) and papaya (Ali et al., 2014)
after ozone treatment. The increase in the ascorbic acid content was related to several reasons such
as inhibition of ascorbate peroxidase and ascorbate oxidase (An et al., 2007) and also ozone stress
may lead to biosynthesis of ascorbic acid by utilizing carbohydrate reserves (Perez et al., 1999).
Yeoh et al. (2014) observed that the vitamin content in fresh-cut papaya was increased up to 20 min
of treatment; however, after 30 min of treatment it decreased significantly when compared to that
of control. It may be attributed to antioxidant property of ascorbic acid content scavenging the
free radicals generated during auto-decomposition of ozone. Similar results were also reported for
phenolic and flavonols contents of banana and fresh-cut honey pineapple (Ananas comosus Merr)
(Alothman et al., 2010).
Polyphenols or total phenolics content was reported to be increased in red bell peppers (Glowacz
et al., 2015), fresh-cut papayas (Yeoh et al., 2014), kiwi fruit (Minas et al., 2012), pineapple, and
banana (Alothman et al., 2010). It may be attributed to the activation of phenylalanine ammo-
nium lyase enzyme, and it is the key enzyme for synthesis of phenolic compounds in plant tissues.
Additionally, inactivation of polyphenol oxidase and peroxidase enzyme may increase the total phe-
nolic content (Chauhan et al., 2011). Swami et al. (2016) assessed the effect of ozone treatment on
11 polyphenols in apple. They observed that the overall concentration of polyphenols was increased
after ozone treatment but individually polyphenols showed variable results. However, total phenolic
content of guava was decreased with the treatment time (Alothman et al., 2010).
Anthocyanins and flavonols were found to decrease in raspberries, strawberries, and blueberries
after ozone treatment (Yaseen et al., 2014). It was related to free radical scavenging ability causing
reduction in their concentration. However, Alothman et al. (2010) reported that total flavonoid content
(TFC) of banana and pineapple increased up to 20 min of exposure to ozone gas and decreased thereaf-
ter up to 30 min. However, in case of guava fruit TFC increased up to 10 min and decreased afterwards.
The fructose and glucose were found to increase in red bell peppers exposed to 0.1 μmol mol−1
ozone compared to control; however, peppers exposed to 0.3 μmol mol−1 of ozone showed no
changes (Glowacz et al., 2015). The vegetables such as cucumber and zucchini showed no biochemi-
cal changes which was attributed to limited penetration through the cuticle (Glowacz et al., 2015).
11.6.1.4 Effect on Pesticide Residue
Pesticides are used to protect the fruits and vegetables from pests and diseases which affect their
natural growth. However, use of non-recommended and higher dose of pesticides leaves a residue
in fruits and vegetables after harvesting (Swami et al., 2016). These trace amounts of pesticide
residue lying in the human body may turn into carcinogen can cause cancer (Carrozza et al., 2009).
The ozonation, apart from antimicrobial activity, also has the ability to remove the pesticides by
40%–99%, which has been achieved successfully in several fruits and vegetables (Whangchai et al.,
2006; Kusvuran et al., 2012; Chen et al., 2013; Upadhyay et al., 2014).
Effect of aqueous ozone on detoxification of six pesticides, i.e., chlorpyrifos, cypermethrin,
azoxystrobin, hexaconazole, methyl parathion, and chlorothalonil from apple fruits was studied
(Swami et al., 2016). Ozonation for 15 min decreased residues of the test pesticides in the range
of 26.91%–73.58%, while ozonation for 30 min could remove the pesticide residues by 39.39%–
95.14%. Kusvuran et al. (2012) reported that the ozone was used to remove the pesticides such as
chlorpyrifos ethyl, tetradifon, and chlorothalonil from lemon, orange, and grape fruit matrices.
It was observed that all the pesticides removed completely. Additionally, increasing temperature
during ozone application showed negative effect and increasing ozone concentration has showed
non-significant changes in the pesticides residues. Tabakoglu and Karaca (2015) studied the effect of
ozone (1.0 ppm) on degradation of azoxystrobin, a fungicide used in grapes. A 90% ± 1% of degrada-
tion of fungicide was obtained after 3 weeks of storage. Kiris and Velioglu (2016) reported that the
treating of olives with ozonated water can remove pesticides by 38%, 50%, 55%, and 61% for chlor-
pyrifos, β-cyfluthrin, α-cypermethrin, and imidacloprid, respectively, after 5 min of ozonated water
washing. However, their levels were higher than national maximum residue levels (MRLs). It may
be attributed to the high oil content, thick surface of the olives, as well as the high pesticide residue
on the olives. The removal efficiency also depends on the structural properties of the pesticides and
matrices. Ikeura et al. (2011) studied the effect of type of ozone microbubble generator (decompres-
sion and gas-water circulation type) and ozone (2 ppm) treatment time (5, 10 min) to remove the
fenitrothion pesticide residue from lettuce, cherry tomatoes, and strawberries. The decompression
type generator was found to be more effective in removal of pesticide than the other one. Ozone at
10 min treatment time effectively reduced the fenitrothion than at 5 min of treatment; however, the
fenitrothion content in lettuce was reduced effectively than in cherry tomatoes and strawberries.
Among cherry tomatoes and strawberries, reduction of fenitrothion was more in strawberries, the
dissolved ozone and hydroxyl radicals could not penetrate through thick pericarp of the cherry toma-
toes and reach the sarcocarp and were inactivated by contact with the pericarp. Chen et al. (2013) also
reported the use of ozone (250 mg/h and 500 mg/h for 15–30 min.) for removal of pesticides such
as permethrin, chlorfluazuron, and chlorothalonil from vegetables, viz. Chinese white cabbage, and
green-stem bok choy. A removal efficiency of 75%–77% was achieved in 15 min treatment time when
ozonation was combined with circulation.
11.6.2 Fruit Juices
Fruit juices are highly susceptible to outbreaks of foodborne pathogens associated with immuno-
compromised results in human beings. These outbreaks led the USFDA to issue hazard analysis and
critical control point (HACCP) regulations for safe and sanitary processing of juice (USFDA, 2001).
Their primary performance standard is a minimum 5-log reduction of the pathogens of concern in
the juices being processed (USFDA, 2001).To date, fruit juices are processed by widely adopted
conventional thermal processing methods to extend their shelf life.
Application of ozone as an alternative processing method has been successfully employed by
several researchers on fruit juices such as apple cider and orange juice (Williams et al., 2004; Tiwari
et al., 2008; Patil et al., 2009; Patil et al., 2010), apple juice (Steenstrup and Floros, 2004; Torres
et al., 2011; Torlak, 2014), blackberry juice (Tiwari et al., 2009), grape juice (Tiwari et al., 2009),
and prebiotic orange juice (Almeida et al., 2015). Steenstrup and Floros (2004) reported that the
overall inactivation of E. coli O157:H7 by ozone is fast enough for practical application in apple
juice production. Torlak (2014) reported that ozone processing can reduce the spore-forming organ-
isms, i.e., Alicyclobacillus acidoterrestris count by 2 logs. However, Torres et al. (2011) reported
that the ozone processing of apple juice reduced the microbial counts by 5 logs, but, meanwhile it
affected the nutrients such as chlorogenic acid, caffeic acid, cinnamic acid, and total phenol content
adversely by 99.1%, 96.6%, 99.8%, and 49.7% of reductions, respectively. Similarly, Patil et al. (2010)
reported that 66.5%, 73.5%, and 65% reduction for chlorogenic acid, caffeic acid, and cinnamic acid,
respectively, was found during ozone processing of orange juice. It was reported that the reductions
in polyphenols were most likely due to the oxidation potential of the ozone. The degradation of
polyphenols may undergo direct reaction with ozone or with their decomposed products. Hence,
while considering ozone as a food-preservation technique, the associated changes in the nutritional
value should also be considered. Similarly, Tiwari et al. (2009) reported that the anthocyanins con-
tent of the blackberry juice decreased with increase in ozone concentration and treatment time.
Meanwhile, the instrumental color values L (brightness) and b (yellowness) values were increased
and a (redness) value decreased with increased ozone concentration and treatment time. Colour
degradation in orange juice due to ozone treatment was observed by Tiwari et al. (2008), who
reported that loss in color is due to the oxidation at conjugated double bonds leading to cleavage
of chromophore of carotenoid, which is responsible for orange juice color. These pigments contain
one or more aromatic rings, the ozone and its decomposition products are responsible for opening
of the aromatic ring and lead to partial oxidation of products such as organic acids, aldehydes, and
ketones. However, Almeida et al. (2015) reported that ozone treatment in prebiotic orange juice did
not affect the phenolic and oligosaccharide content.
11.6.3 Grain Storage
The growing concern over pesticide use and its associated severe health problems in human
beings begins the search for new harmless fumigants in food grains. Registered fumigants used to
eliminate pests during food grain storage include methyl bromide, phosphine, and aluminum phos-
phide (Kells et al., 2001; Tiwari et al., 2010). However, methyl bromide was eliminated by the US
government for use as a fumigant in 2005, due to its adverse effect on the environment (Kells et al.,
2001). The greater risk arises when insects or pests develop resistance to existing fumigants. Apart
from that, another major problem in stored grains is mycotoxin produced by fungal contaminants
due to higher grain moisture content resulting from condensation and leaks (Kells et al., 2001).
Several works have been carried out to evaluate the ability of ozone to reduce the pest or insect
problems, fungal contaminants, and their mycotoxins in food grains.
11.6.3.1 Effect on Pests or Insects
Insect infestation is one of the major concerns in the stored-food grains industry. Improper
control conditions and resistance developed by insects to currently used insecticides are the key
factors for search of novel control methods. Several reports have indicated that ozone has potential
insect killing ability during storage of several food grains without affecting their natural character-
istics (Kells et al., 2001; Leesch, 2003; McDonough et al., 2011). However, the insecticidal effect
of ozone is dependent on several factors such as insect taxonomic families, species, and different
life cycle stages of insects. Among taxonomic families, moths were found more sensitive to ozone
than beetles (Leesch, 2003). McDonough et al. (2011) observed that Plodia interpunctella had 100%
mortality at 500 ppm of ozone when exposed for 60 min. The same concentration and treatment
time resulted only 12% mortality of Tribolium constaneum. However, by using higher concentra-
tion of ozone and longer treatment time can cause 100% mortality of T. constaneum. Isikber and
Oztekin (2009) observed similar differences between two other species from these insect families:
Ephestiakuehniella (Zeller) and Triboliumconfusum adults, belonging to the family Pyralidae and
Tenebrionidae, respectively, when treated with a continuous ozone flow of 13.9 mg/L (6482 ppm)
for 2 h. The adults, pupae, larvae, and eggs of the Ephestiakuehniella showed higher mortality than
Tribolium confusum. Kells et al. (2001) reported that stored maize treated with ozone of 50 ppm for
3 days showed 94.5%, 100%, and 92.2% mortality of Indian meal moth larvae, maize weevil adult,
and red flour beetle adult, respectively. However, the percent of mortality varied with concentra-
tion of ozone used and treatment time. McDonough et al. (2011) observed the differences in ozone
sensitivity at species level, where the Sitophilus oryzae (100% mortality) showed more sensitive to
ozone (at 1800 ppm for 60 min) than Sitophilus zeamais (93% mortality). The different life stages of
insects are egg, pupae, larvae, and adult. Among them eggs are most insensitive to ozone (Isikber
and Oztekin, 2009). The mortality is also first observed in adults followed by larvae, pupae, and
eggs (Leesch, 2003). In early life stages of insects, the lack of respiration is attributed to the resis-
tance towards ozone (Hoback and Stanley, 2001). The respiration undergoes oxidative damage.
However, McDonough et al. (2011) reported that 100% mortality of eggs can be achieved by using
high concentration of ozone (1800 ppm) treatment for long time (180 min).
11.6.3.2 Effect on Microorganisms
Several food grains have been treated with ozone to inactivate the microbial contamination.
Ozone concentration of 50 ppm exposed for 3 days showed 63% reduction in Aspergillus parasiticus
conidia on grain surface (Kells et al., 2001). A. flavus and A. parasiticus are potential aflatoxigenic
species, found both in soil and in the air, which can infect the kernels during pre- and post-harvest
periods. Ozone treatment of 21 ppm for 96 h on peanuts showed 3-log reduction in A. flavus and
A. parasiticus (De Alencar et al., 2012). A. flavus predominantly produces aflatoxins B1 and B2;
A. parasiticus is able to additionally synthesis aflatoxins G1 and G2. Application of ozone in bar-
ley, peanut, and wheat grains showed a significant reduction in major mycotoxins producers such
as Aspergillus and Penicillium species when exposed for 3 min (Ciccarese et al., 2007). Savi et al.
(2014) reported on the complete inhibition of artificially contaminated Fusarium graminearum on
wheat grains. However, the quantity of grains used for ozone treatment is the major point. The treat-
ment of grains by ozone has been divided into two phases. In Phase 1, during initial fumigation, the
inherent sites present on the food grains react with ozone and ozone get degraded. Hence, it takes
more time for fumigation in phase 1 and also as the quantity of grains increases the time of exposure
also increases. In phase 2, once these sites have reacted with ozone, the rate of ozone degradation
decreases (Kells et al., 2001; Mendez et al., 2003).
11.6.3.3 Effect on Mycotoxins
Mycotoxin is a toxic secondary metabolite produced by the organisms of fungus kingdom and
it is capable of causing disease and death in both animals and humans. Food grains are susceptible
to contamination with aflatoxins producing organism, predominantly A. flavus and A. parasiticus,
during harvesting and storage. The ozonation process is also proposed as a technology capable of
degrading mycotoxins, including aflatoxins, fumonisins, ochratoxin, patulin, deoxynivalenol, and
zearalenone. McKenzie et al. (1997) reported that aflatoxin B1 and G1 are most rapidly degraded
with ozonation while aflatoxin B2 and G2 are resistance to oxidation and required higher levels of
O3 for rapid degradation. The difference in degradation rates are due to presence of C8-C9 double
bonds in AfB1 and AfG1, but it is lacking in AfB2 and AfG2. The dipole nature of O3 undergoes
1,3-cycloaddition of O3 at the C8-C9 double bonds. Further, it forms ozonide, which rearrange
into a molozonide derivative, yielding a variety of carbonyl compounds (aldehydes and ketones) or
organic acids in protic solvents. De Alencar et al. (2012) observed that ozone treatment can reduce
the total aflatoxins and aflatoxin B by 30% and 25% in peanuts. However, in another study on ozona-
tion of peanuts showed AfB1 was degraded by 70%, it may be due to the reason that peanuts were
artificially contaminated and, thus, it was found only on the surface (Proctor et al., 2004). Savi
et al. (2014) also observed that the complete depletion of artificially contaminated deoxynivalenol
(mycotoxin) from pericarp and endosperm of wheat grain. The mycotoxin which is produced by
fungus may be distributed throughout the cotyledon and it is protected by grain outer layers; hence,
it is often considered difficult to remove from the grains (De Alencar et al., 2012). However, in real
practical situations, application of ozone is affected by the nature of food products. The ozone is
self-degraded when reacted with food materials causing reduction in their effectiveness against
microorganisms.
It has been proved that application of ozone is having strong remedial activity on insects/pests,
microorganisms, and their toxins. However, the use of high concentration of ozone may also involve
in affecting the nutritional and functional quality of the grains. It was reported that treatment of
grains with 50 ppm ozone for 30 days had no detrimental effect on popping volume of popcorn;
fatty acid and amino acid composition of soybean, wheat, and maize; milling characteristics of
wheat and maize; baking characteristics of wheat; and stickiness of rice (Mendez et al., 2003).
Similar results were also obtained for the wheat grains treated with ozone concentration of 40 and
60 μmol/mol exposed up to 180 min showing no changes in lipid peroxidation and total protein pro-
file (Savi et al., 2014). From the above studies, it can be concluded that the food grains have greater
protection for their nutrients against ozone action.
11.6.4 Meat
Meat is animal flesh that is eaten as food. It is composed of majorly fat and protein. Generally,
meat is highly prone to microbial contamination due to its rich nutrients, high moisture content, and
higher pH. This favorable environment leads to growth of spoilage and pathogenic bacteria inducing
off-flavor, discoloration, and production of toxins. Ozonation is one of the surface decontamination
processes that could be applied to increase the microbial stability of meat and meat products.
The ozone showed antimicrobial effect over number of microorganisms in meat and meat prod-
ucts such as E. coli, Salmonella, Listeria, and Clostridium species. The oxidizing effect of ozone
is a major principle mechanism which affects cell wall and cell membrane leading to cell lysis and
eventually inactivating microorganisms. Also, the ozone processing leaves no residues and no pro-
duction of carcinogens like chlorine-based sanitizers used commonly in meat processing (Novak
and Yuan, 2003).
The beef, mutton, and chevon may get contaminated during fabrication reducing the shelf life
of the meat products. However, immediate washing treatments will reduce the microbial load and
also avoid the spreading of microorganisms to newly cut/exposed surfaces. Application of ozone has
been tried to decontaminate the fabricated meat surfaces and to avoid contaminants during process-
ing and storage of the meat. Stivarius et al. (2002) reported that the beef treated with 1% ozone water
for 7 and 15 min affected the E. coli, Salmonella typhimurium, coliforms, and aerobic bacteria
significantly. However, exposure period of 7 minutes affected only Salmonella typhimurium and
aerobic bacterial counts, so treatment time is crucial for decontamination. Cho et al. (2014) reported
that the ground Hanwoo beef artificially inoculated with E. coli when exposed continuously to
ozone (10 × 10−6 at 4°C for 3 days) during storage caused significant reduction in microbial counts.
However, the above studies have highlighted that the ozone treatment can achieve only limited
microbial reductions on meat surfaces. This may be due to the use of sub-lethal ozone concentra-
tion during treatment of meat surface. The use of higher ozone concentration to achieve increased
inactivation of pathogens may indulge in off-flavour generation and discoloration (Rasanayagam,
2006). Rasanayagam (2006) used 10,000 ppm of ozone to treat beef and poultry inoculated with
Salmonella and the treatment showed 2-log reduction in the microbial count. McMillin and Michel
(2000) used 500, 3500, and 5000 ppm of ozone which caused 2-log reduction in E. coli counts, and
the efficacy increased with increase in the concentration of ozone. However, Cardenas et al. (2011)
reported that the ozone concentration of 72 ppm (low ozone concentration) exposed for 24 h at 0°C
caused 0.7- and 2-log reduction in E. coli and total aerobic mesophilic heterotrophic microorgan-
isms counts, respectively. However, high reactivity of ozone may also involve in reaction with food
organic components; hence, their antimicrobial activity gets reduced. Ozone technology, therefore,
can be used to complement the current food processing practices. In this way, Novak and Yuan
(2003) artificially inoculated beef with Clostridium perfringens, E. coli, and Listeria monocyto-
genes and treated with ozone (3 ppm for 5 min) in combination with heat, sodium salt (NaCl),
and pH individually; it was found that ozone treatment alone did not sterilize the meat effectively.
However, ozone treatment decreased the resistance of vegetative cells of Clostridium perfringens
over subsequent heating at 60°C. Similar results were obtained for NaCl and pH treatments fol-
lowed by ozone processing. With increase in pH, the inactivation was also increased; however, pH
above 12 did not show any effect. Similarly, increased in NaCl concentration increased the inacti-
vation of microorganisms. Pohlman et al. (2002) reported that beef, when treated with 1% ozone
water followed by 5% acetic acid and 0.5% cetylpyridinium chloride, significantly reduced E. coli,
Salmonella typhimurium, coliforms, and aerobic plate count during storage.
Poultry meats are highly perishable due to bacterial contamination; therefore, elimination of
bacterial contaminants is an essential step to avoid loss of quality (Muhlisin et al., 2015). It has
been observed that ozonation (10 ppm) can retain freshness of chicken legs packed in polyamide/
polyethylene bags stored for 12 days at 4 ± 1°C, and shelf life was increased by 4 days compared
to control. It was also reported that ozone-treated chicken legs showed decrease in yeasts, molds,
enterobacteriaceae, and lactic acid bacteria; however, total viable count and Pseudomonas species
counts were increased during storage (Gertzou et al., 2016). Muhlisin et al. (2016) reported that
continuous exposure of ozone concentration of 10 × 10−6 kg O3/m3/h during storage of duck and
chicken breast fillets at 4°C ± 1°C for 4 days showed significant decrease in the counts of coliforms,
aerobic, and anaerobic bacteria. Same authors conducted similar experiments, where chicken breast
was inoculated with Salmonella typhimurium and treated with ozone (10 × 10−6 kg O3/m3/h for
3 days), and the ozone treatment significantly reduced the Salmonella typhimurium, aerobic, and
anaerobic bacteria counts (Muhlisin et al., 2015). Muthukumar and Muthuchamy (2013) reported
that raw chicken samples inoculated with Listeria monocytogenes and exposed to ozone of 33 mg/
min till 9 min, showed inactivation up to 2 × 106 CFU/g. the results clearly revealed that the ozone
treatments could be applied to raw chicken samples before they reach outlets for consumers.
The major principle of ozone action is oxidative reactions lead by them during inactivation of
microorganisms. However, oxidative reactions cause deteriorative reactions in functional, sensory,
and nutritional quality of meat and meat products (Cardenas et al., 2011). Meat contains high num-
ber of pro-oxidants (iron and myoglobin in blood) and polyunsaturated fatty acids present in muscle
tissues. Loss of iron, myoglobin, and oxymyoglobin (pigment) during oxidation leads to loss of red
color of meat (discoloration) and oxidation of unsaturated fat produces off-flavors. The beef samples
treated with gaseous ozone (154 × 10−6 kg.m−3 for 24 h at 0 C and 4°C) showed decrease in instru-
mental a* (redness) color value indicating loss of myoglobin and oxymyoglobin which get converted
into metmyoglobin (Cardenas et al., 2011). Meanwhile, the thiobarbituric acid-reactive substance
(TBARS) also increased after 24 h, indicating oxidation of fat. However, ozone treatment up to 3 h
did not show any changes in oxidation. Lipid oxidation (TBARS value) was higher at the treatment
temperature of 0°C compared to 4°C. It was concluded that as the temperature increases disso-
ciation of ozone also increases; hence, at lower temperature (0°C), maximum amount of dissolved
ozone is present as such in muscle tissues. Similar results were also obtained for Hanwoo beef
treated with gaseous ozone (10 × 10−6 kg O3 h−1) showing increase in TBARS values and decrease
in a* values, whereas, catalase and glutathione enzyme were not affected by ozone treatment (Cho
et al., 2014). However, Stivarius et al. (2002) observed increase in lightness (L*) values only in beef
after treatment with 1% ozone water for 7 and 15 min, and no significant change in redness (a*)
and sensory values were observed. Muhlisin et al. (2016) observed that ozonation induces faster
oxidation (TBARS) of duck breast fillet than control and chicken breast fillets; it may be due to the
high unsaturated fatty acid content in duck breast fillet compared to chicken breast fillet. Similarly,
ozonation reduced the activity of catalase in duck breast fillets significantly; however, its activity
was not affected in chicken breast fillets. Ozonation also reduced the activity of glutathione peroxi-
dase in both the samples, but they were found to be less sensitive to ozone as compared to catalase.
It was concluded that because of decrease in activity of these enzymes increased lipid oxidation
rates. Eventually, the redness (a*) of the both samples also reduced after ozonation and also during
storage, higher declining rate was found to be more in duck breast fillets. Meanwhile, the L*, b*,
and h° were increased on surface of duck and chicken breast fillets. It may be attributed to oxidation
of myoglobin and oxymyoglobin into metmyoglobin. Similar results were obtained for the chicken
breast when continuously treated with ozone during storage. Ozonation increases TBARS values,
inhibit the catalase and glutathione peroxidase, increases b* values, and decreases the L* and a*
values (Muhlisin et al., 2015).
11.6.5 Fish
Fish is a highly perishable food, contaminated heavily with spoilage microorganisms. Therefore,
it has been restricted to consume fresh once it has been lured. Bacterial contamination leads to deg-
radation of fish constituents, particularly non-protein nitrogenous compounds, which gives typical
fish spoilage odour. Usually fishing is done for long times (15–17 days) in deep sea, so quality may
get worsen. In addition, the fresh quality is desired by consumers. Traditionally, fish is washed after
caught using sea water to remove the slime on the fish surface, which is a source of contamination
and also stored at low temperature using ice made up of sea water. However, several incidents showed
the traditional system compromised safety and shelf life of the fish when stored for a long time. So,
to carry fish safe and fresh for long time till it reaches to consumers, the handling and storage condi-
tions are key parameters which affect the shelf life and quality. However, it should always be kept
in mind that the technological interventions intended to enhance the shelf life should not affect the
original sensory qualities of the fish. In this context, the treatments which are given to fish should
keep fish safe without affecting its original quality and it should not leave any harmful residues
after processing. Ozonation as a decontamination process attracts in processing of several foods
including several varieties of fish. The European directive for fish products recommends the total
count of mesophilic aerobic bacteria as indicating bacteria for fish deterioration in the limit of 5–6
logarithmic units. Pastoriza et al. (2008) reported use of ozonated water (2 ppm) for washing of fish
(Hake, Merluccius merluccius) and to manufacture ice in flakes (Petfrost system) against tradition-
ally used sea water (control). The fish washed and stored in ozonated water ice showed better stability
against total aerobic microorganisms compared to control. Temperature also plays a major role dur-
ing ozone treatment, which was also proved when fish treated at 0°C showed bactericidal action,
against 5°C (Gelman et al., 2005). Da Silva et al. (1998) reported that ozonation showed bactericidal
action against total viable counts, enterobacteriaceae, obligate psychrophiles, pseudomonadaceae,
bacterial H2S producers, and lactic acid bacteria present on skin of fresh scad (Trachurus trachurus)
by 1-log reduction by using ozone (<0.27 × 10−3 gL−1 for 10 days). Whereas, ozonation of fish muscle
achieved only 1-log reduction of total viable counts and H2S producers. It may be due to less acces-
sibility of microorganisms for ozone action when present in the muscles compared to surface. Lu
et al. (2012) observed slow growth in total viable count in sea bass (Lateolabrax japonicus) treated
with ozonized water and stored in ozonized ice flakes for 18 days. Chawla et al. (2007) optimized
the ozone concentration and treatment time of 3 ppm for 60 s soaking for peeled shrimp meat for
inactivating total aerobic bacteria and pseudomonas species. They compared the soaking treatment
with spraying treatment, the former found to be more effective in inactivating bacteria. However, it
has been found that spraying treatment is effective in dealing with high-fat fish varieties to balance
both microbial safety and chemical quality. The spraying method reduces the contact time between
fish and ozone, extent of oxidation of fat may reduce. Crowe et al. (2012) studied the effect of spray-
ing application of ozonated water (1 and 1.5 ppm) over high-fat contained salmon fillets followed by
storage for 10 days at 4°C. The study reported that ozone treatment reduces Listeria monocytogenes
content effectively during storage. Nerantzaki et al. (2005) reported that rainbow trout ozonized,
vacuum packed, and stored at 4 ± 0.5°C for 15 days. The ozone treatment affected the population of
mesophilic aerobic bacteria, Pseudomonas species, and H2S-producing bacteria until day 11 of stor-
age, Brochothrix thermosphacta, lactic acid bacteria, and Enterobacteriaceae until day 8 of stor-
age. Similar results obtained for sardine (Sardina pilchardus) stored in ozonized slurry ice showing
a shelf life of 19 days as compared to samples stored in slurry ice or flaked ice with shelf lives of
15 and 8 days, respectively (Campos et al., 2005). Chen et al. (2014) reported that shucked oysters
treated with ozonated water (9 ppm for 10 min at 5°C) significantly reduced the total aerobic plate
counts by 2 logs. Tantratian et al. (2011) reported that ozonated water (220–1130 ppm) when used
for treating black tiger prawn (Penaeus monodon), the inactivation rate also increased with increase
in ozone concentration. The E. coli and Vibrio parahaemolyticus were found to be more sensitive to
ozone than Salmonella anatum. Okpala (2014) stored white shrimp under minimal ozone treatment
for 11 days. The aerobic plate count reduced during early storage period significantly compared to
control, and it increased at later stages of storage. Ozonation of shucked mussels done at the concen-
tration of 1 ppm for 60 and 90 min decreased the microbial populations such as aerobic plate count,
Pseudomonas species, H2S producing bacteria, Brochothrix thermosphacta, Lactic acid bacteria,
and Enterobacteriaceae. The shelf life was increased by 3 days compared to control (Manousaridis
et al., 2005).
Fishes are good source of high-quality protein and fat. Therefore, peeling, cutting, etc. of fish
during ozone processing cause some loss in nutritional value and also increases the off-odour due
to oxidation of fat. Ozone participates in direct or indirect oxidation by ozonolysis and catalysis.
Through oxidation property, it oxidizes polypeptide backbone of protein, peptide bond cleavage,
protein cross-linking, and several amino acids modification. This structural modification alters the
functional property of proteins. So oxidative damage created by ozone not only affects fat or lipid but
also creates oxidative stress on protein, associated with chemical deterioration of foods. Oxidation
of proteins increases the carbonyl groups, degradation of essential amino acids, loss of sulfhydryl
groups, and formation of intra/inter molecular crosslinking. Zhang et al. (2016) studied the effect
of ozonation on protein and fat constituents in the meat of bighead carp (Hypophthalamichthys
nobilis). It was reported that the treatment increased salt-extractable protein content, Ca2+ ATPase
activity, total sulfhydryl and active sulfhydryl content, carbonyl content of myofibrillar protein and
gel strength without affecting peroxide value, and thiobarbituric acid value, which are related to
oxidation of lipids. The ozone treatment did not show any changes in protein distribution and new
patterns on SDS PAGE. This indicates that ozone induces oxidation of protein, where proteins
break into smaller fragments without formation of new fragments through crosslinking. Dehkordi
and Zokaie (2010) observed that ozonation increased the shelf life of fish by 2 days by delaying the
peroxide value and total volatile nitrogen. Pastoriza et al. (2008) reported that total volatile bases
(TBA) and trimethylamine content (TMA) of a treated sample of fresh fish (Hake, Merluccius
merluccius), when washed with ozonised water and stored in ozonized ice for 18 days, were found
to be below the legal limits even after 17 days of storage. According to the European Commission,
the levels of TBA of 30 mg per 100 g of fish muscle can be considered as deteriorated and unsuit-
able for consumption. TMA is often considered as good quality indicator of fish products stored in
refrigeration. TMA is product of bacterial action during refrigerated storage, its level of 10–15 mg
per 100 g were suggested as maximum limit for acceptability of fresh fish (Connell, 1995). Similar
results reported for seabass (Lu et al., 2012), peeled shrimp meat (Chawla et al., 2007), rainbow
trout (Nerantzaki et al., 2005), salmon fillets (Crowe et al., 2012), Sardine (Campos et al., 2005),
shucked oysters (Chen et al., 2014), black tiger prawn (Tantratian et al., 2011), and white shrimp
(Okpala, 2014).
11.7 SaFEtY aND rISKS
One has to take care while dealing with ozone gas as ozone is a toxic gas, and continuous
inhalation can cause severe illness and even death. It is highly oxidant and therefore shows
strong toxicity to animals, plants, and living organisms. Toxicity symptoms, such as sharp
irritation to the nose and throat could, result instantly at 0.1 ppm dose. Loss of vision could
arise from 0.1 to 0.5 ppm after exposure for 3–6 h. Ozone toxicity of 1–2 ppm could cause
distinct irritation on the upperpart of throat, headache, pain in the chest, cough, and drying of
the throat. Higher levels of ozone (5–10 ppm) could cause increase pulse, and oedema of lungs.
Ozone level of 50 ppm or more is potentially fatal (Muthukumarappan et al., 2000). Ozone is a
toxic gas; toxicity is dependent on concentration and length of exposure (Pascual et al., 2007).
At short-term exposure rates of 0.1–1.0 ppm, symptoms include headaches, nosebleeds, eye irri-
tation, dry throat, and respiratory irritation. At higher exposure levels (1–100 ppm), symptoms
become more severe and include asthma-like symptoms, tiredness, and loss of appetite (Greene
et al., 2012).
rEFErENCES
Ali, A., Ong, M. K., and Forney, C. F. (2014). Effect of ozone pre-conditioning on quality and antioxidant
capacity of papaya fruit during ambient storage. Food Chem. 142: 19–26.
Al-Haddad, K. S. H., Al-Qassemi, R. A. S., and Robinson, R. K. (2005). The use of gaseous ozone and gas
packaging to control populations of Salmonella infantis and Pseudomonas aeruginosa on the skin of
chicken portions. Food Control 16(5): 405–410.
Almeida, F. D. L., Cavalcante, R. S., Cullen, P. J., Frias, J. M., Bourke, P., Fernandes, F. A., and Rodrigues, S.
(2015). Effects of atmospheric cold plasma and ozone on prebiotic orange juice. Innov Food Sci. Emerg
Technol. 32: 127–135.
Alothman, M., Kaur, B., Fazilah, A., Bhat, R., and Karim, A. A. (2010). Ozone-induced changes of antioxidant
capacity of fresh-cut tropical fruits. Innov Food Sci. Emerg Technol. 11(4): 666–671.
Alwi, N. A., and Ali, A. (2014). Reduction of Escherichia coli O157, Listeria monocytogenes and Salmonella
enterica. Typhimurium populations on fresh-cut bell pepper using gaseous ozone. Food Control 46:
304–311.
An, J., Zhang, M., and Lu, Q. (2007). Changes in some quality indexes in fresh-cut green asparagus pretreated
with aqueous ozone and subsequent modified atmosphere packaging. J Food Eng. 78: 340–344.
Bablon, G., Bellamy, W. D., Billen, G., Bourbigot, M. M., Daniel, B., Erb, F., and Gomella, C. et al. (1991).
Practical application of ozone: Principles and case studies. Ozone in Water Treatment: Application and
Engineering 569.
Battino, R., Rettich, T. R., and Tominaga, T. (1983). The solubility of oxygen and ozone in liquids. J Phys
Chem Ref Data 12(2): 163–178.
Blatchley, E. R., and Hunt, N. K. (2002). Ozone disinfection in drinking water. American Society Civil Eng.
2002: 241–271.
Campos, C. A., Rodríguez, O., Losada, V., Aubourg, S. P., and Barros-Velazquez, J. (2005). Effects of storage
in ozonised slurry ice on the sensory and microbial quality of sardine (Sardina pilchardus). Int J Food
Microbiol. 103(2): 121–130.
Cardenas, F. C., Andres, S., Giannuzzi, L., and Zaritzki, N. (2011). Antimicrobial action and effects on beef
quality attributes of a gaseous ozone treatment at refrigeration temperatures. Food Control 22: 1442–1447.
Carrozza, S. E., Li, B., Wang, Q., Horel, S., and Cooper, S. (2009). Agricultural pesticides and risk of child-
hood cancers. Int J Hyg Envir Heal. 212: 186–195.
Chauhan, O. P., Raju, P. S., Ravi, N., Singh, A., and Bawa, A. S. (2011). Effectiveness of ozone in combination
with controlled atmosphere on quality characteristics including lignification of carrot sticks. J Food
Eng. 102(1): 43–48.
Chawla, A., Bell, J. W., and Janes, M. E. (2007). Optimization of ozonated water treatment of wild-caught and
mechanically peeled shrimp meat. J Aquat Food Prod T. 16(2): 41–56.
Chen, H., Wang, M., Chen, S., Chen, T., and Huang, N. (2014). Effects of ozonated water treatment on the
microbial population, quality, and shelf life of shucked oysters (Crassostrea plicatula). J Aquat Food
Prod T. 23(2): 175–185.
Chen, J. Y., Lin, Y. J., and Kuo, W. C. (2013). Pesticide residue removal from vegetables by ozonation. J Food
Eng. 114(3): 404–411.
Cho, Y., Choi, J. H., Hahn, T. W., and Lee, S. K. (2015). Bacterial counts and oxidative properties of chicken
breast inoculated with Salmonella Typhimurium exposed to gaseous ozone. J Food Saf. 35(1): 137–144.
Cho, Y., Muhlisin, J. H. C., Hahn, T. W., and Lee, S. K. (2014). Effect of gaseous ozone exposure on the bacte-
ria counts and oxidative properties of ground hanwoo beef at refrigeration temperature. Korean J Food
Sci Anim Resour. 34(4): 525.
Choi, M. R., Liu, Q., Lee, S. Y., Jin, J. H., Ryu, S., and Kang, D. H. (2012). Inactivation of escherichia coli
O157: H7, salmonella typhimurium and listeria monocytogenes in apple juice with gaseous ozone. Food
Microbial. 32(1): 191–195.
Ciccarese, F., Sasanelli, N., Ciccarese, A., Ziadi, T., and Mancini, L. (2007, October). Seed disinfestation by
ozone treatments. In Proceedings of the IOA Conference and Exhibition Valencia, Spain Europe.
Connell, J. J. (1995). Control of Fish Quality. Osney meat. Oxford, UK: Fishing News Books Ltd.
Crowe, K. M., Skonberg, D., Bushway, A., and Baxter, S. (2012). Application of ozone sprays as a strategy to
improve the microbial safety and quality of salmon fillets. Food Control 25(2): 464–468.
Da Silva, M. V., Gibbs, P. A., and Kirby, R. M. (1998). Sensorial and microbial effects of gaseous ozone on
fresh scad (Trachurus trachurus). J Appl Microbial. 84(5): 802–810.
Dehkordi, B. M., and Zokaie, N. (2010). Extension of fish shelf life by ozone treatment. World Academy Sci
Eng Technol. 62: 150–152.
De Alencar, E. R., Faroni, L. R. D. A., Soares, N. D. F. F., da Silva, W. A., and da Silva Carvalho, M. C. (2012).
Efficacy of ozone as a fungicidal and detoxifying agent of aflatoxins in peanuts. J Sci Food Agric. 92(4):
899–905.
De Smedt, F., De Gendt, S., Heyns, M. M., and Vinckier, C. (2001). The application of ozone in semiconductor
cleaning processes: The solubility issue. J Electrochem Soc. 148(9): G487–G493.
Gao, Y. H., Wang, W. S., Chen, C. K., Dong, C. H., and Li, H. J. (2008). Study on factors of influencing stabil-
ity of high-concentration ozonated water [J]. Storage Proces. 6: 016.
Gelman, A., Sachs, O., Khanin, Y., Drabkin, V., and Glatman, L. (2005). Effect of ozone pretreatment on fish
storage life at low temperatures. J Food Prot. 68: 778–784.
Gertzou, I. N., Drosos, P. E., Karabagias, I. K., and Riganakos, K. A. (2016). Combined effect of ozonation
and packaging on shelf life extension of fresh chicken legs during storage under refrigeration. J Food
Sci Technol. 53(12): 4270–4277.
Glowacz, M., Colgan, R., and Rees, D. (2015). Influence of continuous exposure to gaseous ozone on the qual-
ity of red bell peppers, cucumbers and zucchini. Postharvest Biol Technol. 99: 1–8.
Gonçalves, A. A. (2016). Ozone as a safe and environmentally friendly tool for the seafood industry. J Aquat
Food Prod T. 25(2): 210–229.
Greene, A. K., Guzel-Seydim, Z. B., and Seydim, A. C. (2012). Chemical and physical properties of ozone. In:
Ozone in Food Processing, pp. 19–31. O´Donnell, C., Tiwari, B. K., Cullen, P. J., and Rice, R. G., Eds.,
John Wiley & Sons, New York.
Guzel-Seydim, Z., Bever, P. I., and Greene, A. K. (2004). Efficacy of ozone to reduce bacterial populations in
the presence of food components. Food Microbiol. 21(4): 475–479.
Hill, A. G., and Rice, R. G. (1982). Historical background, properties and applications. In: Handbook of Ozone
Technology and Applications, Vol. 1, pp. 1–37. Rice, R. G., and Netzer, A., Eds. Ann Arbor, MI: Ann
Arbor Science Publishers.
Hoback, W. W., and Stanley, D. W. (2001). Insects in hypoxia. J Insect Physiol. 47(6): 533–542.
Horvath, M., Bilitzky, L., and Huttner, J. (1985). Fields of utilization of ozone. In: Ozone. pp. 257–316. Elsevier
Science Publishing, New York.
Horvitz, S., and Cantalejo, M. J. (2012). Effects of ozone and chlorine postharvest treatments on quality of
fresh‐cut red bell peppers. Int J Food Sci Technol. 47(9): 1935–1943.
Hunt, N. K., and Marinas, B. J. (1999). Inactivation of Escherichia coli with ozone: Chemical and inactivation
kinetics. Water Res. 33(11): 2633–2641.
Ikeura, H., Kobayashi, F., and Tamaki, M. (2011). Removal of residual pesticide, fenitrothion, in vegetables by
using ozone microbubbles generated by different methods. J Food Eng. 103(3): 345–349.
Isikber, A. A., and Oztekin, S. (2009). Comparison of susceptibility of two stored-product insects, Ephestia-
kuehniella Zeller and Triboliumconfusum du Val to gaseous ozone. J Stored Prod Res. 45(3): 159–164.
Karaca, H., and Velioglu, Y. S. (2007). Ozone applications in fruit and vegetable processing. Food Reviews
Int. 23(1): 91–106.
Kells, S. A., Mason, L. J., Maier, D. E., and Woloshuk, C. P. (2001). Efficacy and fumigation characteristics of
ozone in stored maize. J Stored Prod Res. 37(4): 371–382.
Ketteringham, L., Gausseres, R., James, S. J., and James, C. (2006). Application of aqueous ozone for treating
pre-cut green peppers (Capsicum annuum L.). J Food Eng. 76(1): 104–111.
Khadre, M. A., Yousef, A. E., and Kim, J. G. (2001). Microbiological aspects of ozone applications in food:
A review. J Food Sci. 66(9): 1242–1252.
Kim, J. G., Yousef, A. E., and Chism, G. W. (1999). Use of ozone to inactivate microorganisms on lettuce,
J Food Safety 19(1): 17–34.
Kim, J. G., Yousef, A. E., and Khadre, M. A. (2003). Ozone and its current and future application in the food
industry. Adv Food Nutr Res. 45: 167–218.
Kiriş, S., and Velioglu, Y. S. (2016). Reduction in pesticide residue levels in olives by ozonated and tap water
treatments and their transfer into olive oil. Food Addit Contam: Part A 33(1): 128–136.
Kusvuran, E., Yildirim, D., Mavruk, F., and Ceyhan, M. (2012). Removal of chloropyrifos ethyl, tetradifon and
chlorothalonil pesticide residues from citrus by using ozone. J hazardous Mater. 241: 287–300.
Leesch, J. G. (2003). The mortality of stored-product insects following exposure to gaseous ozone at high
concentrations. In: Advances in Stored Product Protection. pp. 22–26. Credland, P. F., Armitage, D. M.,
Bell, C. H., Cogan, P. M., and Highley. E., Eds. Proceedings of the 8th International Working Conference
on Stored-Product Protection, York, UK.
Levanov, A. V., Kuskov, I. V., Antipenko, E. E., and Lunin, V. V. (2008). The solubility of ozone in aqueous
solutions of sulfuric, phosphoric, and perchloric acids. Russ J Phys Chem A, Focus on Chem. 82(7):
1126–1131.
Lu, F., Liu, S. L., Liu, R., Ding, Y. C., and Ding, Y. T. (2012). Combined effect of ozonized water pretreatment
and ozonized flake ice on maintaining quality of Japanese sea bass (Lateolabrax japonicus). J Aquat
Food Prod T. 21(2): 168–180.
Mahapatra, A. K., Muthukumarappan, K., and Julson, J. L. (2005). Applications of ozone, bacteriocins and
irradiation in food processing: A review. Crit Rev Food Sci Nutr, 45(6), 447–461.
Manousaridis, G., Nerantzaki, A., Paleologos, E. K., Tsiotsias, A., Savvaidis, I. N., and Kontominas, M. G.
(2005). Effect of ozone on microbial, chemical and sensory attributes of shucked mussels. Food
Microbiol. 22(1): 1–9.
McDonough, M. X., Mason, L. J., and Woloshuk, C. P. (2011). Susceptibility of stored product insects to high
concentrations of ozone at different exposure intervals. J Stored Prod Res. 47(4): 306–310.
McKenzie, K. S., Sarr, A. B., Mayura, K., Bailey, R. H., Miller, D. R., Rogers, T. D., and Phillips, T. D. (1997).
Oxidative degradation and detoxification of mycotoxins using a novel source of ozone. Food Chem
Toxicol. 35(8): 807–820.
McMillin, K., and Michel, M. (2000). Reduction of E. coli in ground beef with gaseous ozone. Louisiana
Agriculture 43(3): 35.
Mendez, F., Maier, D. E., Mason, L. J., and Woloshuk, C. P. (2003). Penetration of ozone into columns of
stored grains and effects on chemical composition and processing performance. J Stored Prod Res.
39(1): 33–44.
Minas, I. S., Tanou, G., Belghazi, M., Job, D., Manganaris, G. A., Molassiotis, A., and Vasilakakis, M. (2012).
Physiological and proteomic approaches to address the active role of ozone in kiwifruit post-harvest
ripening. J Exp Bot. 63(7): 2449–2464.
Muhlisin, M., Cho, Y., Choi, J. H., Hawn, T., and Lee, S. K. (2015). Bacterial counts and oxidative properties of
chicken breast inoculated with Salmonella Typhimurium exposed to gaseous ozone. J Food Saf. 35: 137–144.
Muhlisin, M., Utama, D. T., Lee, J. H., Choi, J. H., and Lee, S. K. (2016). Effects of gaseous ozone exposure
on bacterial counts and oxidative properties in chicken and duck breast meat. Korean J Food Sci Anim
Resour. 36(3): 405.
Muthukumar, A., and Muthuchamy, M. (2013). Optimization of ozone in gaseous phase to inactivate Listeria
monocytogenes on raw chicken samples. Food Res Int. 54(1): 1128–1130.
Muthukumarappan, K., Halaweish, F., and Naidu, A. S. (2000). Ozone. In Natural food antimicrobial systems.
pp. 783–800, Naidu, A. S., Ed. CRC Press, New York.
Navarre, W. W., and Schneewind, O. (1999). Surface proteins of gram-positive bacteria and mechanisms of
their targeting to the cell wall envelope. Microbiol Mol Biol Rev. 63(1): 174–229.
Nerantzaki, A., Tsiotsias, A., Paleologos, E. K., Savvaidis, I. N., Bezirtzoglou, E., and Kontominas, M. G.
(2005). Effects of ozonation on microbiological, chemical and sensory attributes of vacuum-packaged
rainbow trout stored at 4 ± 0.5°C. Eur Food Res Technol. 221(5): 675–683.
Novak, J. S., and Yuan, J. T. (2003). Viability of Clostridium perfringens, Escherichia coli, and Listeria
monocytogenes surviving mild heat or aqueous ozone treatment on beef followed by heat, alkali, or salt
stress. J Food Prot. 66(3): 382–389.
Okpala, C. O. R. (2014). Investigation of quality attributes of ice-stored Pacific white shrimp (Litopenaeusvannamei)
as affected by sequential minimal ozone treatment. LWT-Food Sci Technol. 57(2): 538–547.
Palou, L., Smilanick, J. L., Crisosto, C. H., and Mansour, M. 2001. Effect of gaseous ozone exposure on the
development of green and blue molds on cold stored citrus fruit. Plant Dis. 85: 632–638.
Palou, L., Smilanick, J. L., Crisosto, C. H., Mansour, M., and Plaza, P. (2003). Ozone gas penetration and con-
trol of the sporulation of Penicillium digitatum and Penicillium italicum within commercial packages
of oranges during cold storage. Crop Protection, 22(9): 1131–1134.
Pan, G. Y., Chen, C. L., Chang, H. M., and Gratzl, J. S. (1984). Studies on ozone bleaching. I. The effect of
pH, temperature, buffer systems and heavy metal-ions on stability of ozone in aqueous solution. J Wood
Chem Technol. 4(3): 367–387.
Pascual, A., Llorca, I., and Canut, A. (2007). Use of ozone in food industries for reducing the environmental
impact of cleaning and disinfection activities. Trends Food Sci Technol. 18: S29–S35.
Pastoriza, L., Bernardez, M., Sampedro, G., Cabo, M. L., and Herrera, J. J. (2008). Use of sterile and ozonized
water as a strategy to stabilize the quality of stored refrigerated fresh fish. Food Control 19(8): 772–780.
Patil, S., Bourke, P., Frias, J. M., Tiwari, B. K., and Cullen, P. J. (2009). Inactivation of escherichia coli in
orange juice using ozone. Innov. Food Sci Emerg Technol. 10(4): 551–557.
Patil, S., Valdramidis, V. P., Cullen, P. J., Frias, J. M., and Bourke, P. (2010). Ozone inactivation of acid
stressed Listeria monocytogenes and Listeria innocua in orange juice using a bubble column. Food
Control 21(12): 1723–1730.
Perez, A. G., Sanz, C., Rios, J. J., Olias, R., and Olias, J. M. (1999). Effects of ozone treatment on postharvest
strawberry quality. J Agric Food Chem. 47(4): 1652–1656.
Pohlman, F. W. (2012). Ozone in meat processing. Ozone in Food Processing, pp 123–136, O´Donnell, C.,
Tiwari, B. K., Cullen, P. J., and Rice, R. G., Eds. John Wiley & Sons, UK.
Pohlman, F. W., Stivarius, M. R., McElyea, K. S., Johnson, Z. B., and Johnson, M. G. (2002). The effects
of ozone, chlorine dioxide, cetylpyridinium chloride and trisodium phosphate as multiple antimicro-
bial interventions on microbiological, instrumental color, and sensory color and odor characteristics of
ground beef. Meat Sci. 61(3): 307–313.
Proctor, A. D., Ahmedna, M., Kumar, J. V., and Goktepe, I. (2004). Degradation of aflatoxins in peanut
kernels/flour by gaseous ozonation and mild heat treatment. Food Addit Contam. 21(8): 786–793.
Rasanayagam, V. (2006). Optimization ozone application to increase the selective reactivity. Division 15c
Bioengineering poster session, IFT annual meeting.
Restaino, L., Frampton, E. W., Hemphill, J. B., and Palnikar, P. (1995). Efficacy of ozonated water against
various food-related microorganisms. Appl Environ Microbiol. 61(9): 3471–3475.
Rice, R. G., Robson, C. M., Miller, G. W., and Hill, A. G. (1981). Uses of ozone in drinking water treatment.
J Am Water Works Assoc. 73(1): 44–57.
Rice, R. G. (1986). Application of ozone in water and wastewater treatment. In: Analytical Aspects of Ozone
Treatment of Water and Wastewater, pp. 7–26. Rice, R. G., Bollyky L. J., and Lacy, W. J., Eds. Chelsea,
MI: Lewis Publishers.
Savi, G. D., Piacentini, K. C., Bittencourt, K. O., and Scussel, V. M. (2014). Ozone treatment efficiency on
Fusarium graminearum and deoxynivalenol degradation and its effects on whole wheat grains (Triticum
aestivum L.) quality and germination. J Stored Prod Res. 59: 245–253.
Song, W. J., Shin, J. Y., Ryu, S., and Kang, D. H. (2015). Inactivation of Escherichia coli O157: H7, Salmonella
Typhimurium and Listeria monocytogenes in apple juice at different pH levels by gaseous ozone treat-
ment. J Appl Microbiol. 119(2): 465–474.
Steenstrup, L. D., and Floros, J. D. (2004). Inactivation of E. coli 0157: H7 in apple cider by ozone at various
temperatures and concentrations. J Food Process Preserv. 28(2): 103–116.
Stivarius, M. R., Pohlman, F. W., McElyea, K. S., and Apple, J. K. (2002). Microbial, instrumental color and
sensory color and odor characteristics of ground beef produced from beef trimmings treated with ozone
or chlorine dioxide. Meat Sci. 60(3): 299–305.
Swami, S., Muzammil, R., Saha, S., Shabeer, A., Oulkar, D., Banerjee, K., and Singh, S. B. (2016). Evaluation
of ozonation technique for pesticide residue removal and its effect on ascorbic acid, cyanidin-3-glucoside,
and polyphenols in apple (Malus domesticus) fruits. Environ Monit Assess. 188(5): 1–11.
Tabakoglu, N., and Karaca, H. (2015). Enhanced degradation of azoxystrobin in grapes and model systems by
ozone fumigation during storage. Ozone: Sci Eng. 37(6): 479–488.
Tantratian, S., Setjintanin, J., and Sanguandeekul, R. (2011). Improving the quality of cold storage black
tiger prawn (Penaeus monodon) using ozonated water. In Proceedings of International Conference on
Biotechnology and Food Science, 7th vol. pp 105–109.
Tiwari, B. K., Brennan, C. S., Curran, T., Gallagher, E., Cullen, P. J., and O’Donnell, C. P. (2010). Application
of ozone in grain processing. J Cereal Sci. 51(3): 248–255.
Tiwari, B. K., Muthukumarappan, K., O’Donnell, C. P., and Cullen, P. J. (2008). Kinetics of freshly squeezed
orange juice quality changes during ozone processing. J Agric Food Chem. 56(15): 6416–6422.
Tiwari, B. K., O’Donnell, C. P., Muthukumarappan, K., and Cullen, P. J. (2009). Anthocyanin and colour deg-
radation in ozone treated blackberry juice. Inno Food Sci Emerg Technol. 10(1): 70–75.
Taylor, P. A., Futrell, T. O., Dunn Jr, N. M., Jusko, M. P., DuBois, C. R., and Capehart, J. D. (1996). U.S. Patent
No. 5,547,644. Washington, DC: U.S. Patent and Trademark Office.
Torlak, E., Sert, D., and Ulca, P. (2013). Efficacy of gaseous ozone against Salmonella and microbial popula-
tion on dried oregano. Int J Food Microbiol. 165(3): 276–280.
Torlak, E. (2014). Efficacy of ozone against Alicyclobacillus acidoterrestris spores in apple juice. Int J Food
Microbiol. 172: 1–4.
Torres, B., Tiwari, B. K., Patras, A., Wijngaard, H. H., Brunton, N., Cullen, P. J., and O’Donnell, C. P. (2011).
Effect of ozone processing on the colour, rheological properties and phenolic content of apple juice.
Food Chem. 124(3): 721–726.
Tzortzakis, N., and Chrysargyris, A. (2017). Postharvest ozone application for the preservation of fruits and
vegetables. Food Rev Int. 33(3): 270–315.
United States Food and Drug Administration (USFDA) (2001). United States Food and Drug Administration
(USFDA), hazard analysis and critical point (HACCP); procedures for the safe and sanitary processing
and importing of juice; final rule, Federal Register 66, pp. 6137–6202.
Upadhyay, A., Kalra, S., Saini, P., Niwas, R., Kumar, R., and Gopal, M. (2014). Decontamination of imidaclo-
prid from tomato by ozone treatment and edible alkali. Annals Plant Prot Sci. 22(1): 238–239.
Uzun, H., Ibanoglu, E., Catal, H., and Ibanoglu, S. (2012). Effects of ozone on functional properties of proteins.
Food Chem. 134(2): 647–654.
Varga, L., and Szigeti, J. (2016). Use of ozone in the dairy industry: A review. Int J Dairy Technol. 69(2): 157–168.
Victorin, K. (1992). Review of the genotoxicity of ozone. Mutation Res. 277: 221–238.
Whangchai, K., Saengnil, K., and Uthaibutra, J. (2006). Effect of ozone in combination with some organic acids
on the control of postharvest decay and pericarp browning of longan fruit. Crop Prot. 25(8): 821–825.
Williams, R. C., Sumner, S. S., and Golden, D. A. (2004). Survival of Escherichia coli O157: H7 and sal-
monella in apple cider and orange juice as affected by ozone and treatment temperature. J Food Prot.
67(11): 2381–2386.
Yang, P. P. W., and Chen, T. C. (1979). Stability of ozone and its germicidal properties on poultry meat micro-
organisms in liquid phase. J Food Sci. 44(2): 501–504.
Yaseen, T., Ricelli, R., Albanese, P., Carboni, C., and D’Onghia, A. M. (2014). Effect of storage in ozone-
enriched atmosphere on fungal contamination, catalase, superoxide dismutase and glutathione peroxi-
dase activity in berries. In IOA-EA3G Conference (Dublin, Ireland).
Yeoh, W. K., Ali, A., and Forney, C. F. (2014). Effects of ozone on major antioxidants and microbial popula-
tions of fresh-cut papaya. Postharvest Biol Technol. 89: 56–58.
Zhang, T., Xue, Y., Li, Z., Wang, Y., Yang, W., and Xue, C. (2016). Effects of ozone on the removal of geosmin
and the physicochemical properties of fish meat from bighead carp (Hypophthalmichthys nobilis). Innov
Zuma, F., Lin, J., and Jonnalagadda, S. B. (2009). Ozone-initiated disinfection kinetics of Escherichia coli in
water. J Environ Sci Health, Part A 44(1): 48–56.
ChaptEr 12
Improving the Efficacy of Ozone

treatment in Food preservation
Shyam Ramkrishna Garud, Pradeep Singh Negi, and Navin Kumar Rastogi
CONtENtS
12.1 Introduction ......................................................................................................................... 213

12.2 Ozone Generation Methods ................................................................................................ 215
12.2.1 Corona Discharge Method ..................................................................................... 215
12.2.2 Photochemical Ozone Generator ........................................................................... 216
12.3 Extrinsic Factors ................................................................................................................. 216
12.3.1 Concentration ......................................................................................................... 216
12.3.2 Temperature ........................................................................................................... 217
12.4 Intrinsic Factors .................................................................................................................. 217
12.4.1 pH ........................................................................................................................... 217
12.4.2 Organic Matter Content ......................................................................................... 217
12.5 Ozone as a Preservative ...................................................................................................... 218
12.6 Combination of Ozone with Other Hurdles for Enhancing Preservation Potential ............ 219
12.6.1 Combined Effect of Ozone and Physical Treatment .............................................. 223
12.6.1.1 Ultrasound ............................................................................................. 223
12.6.1.2 Ultraviolet (UV) Radiation....................................................................224
12.6.1.3 Pulsed Electrical Field (PEF) ................................................................ 225
12.6.1.4 Thermal Treatment ................................................................................ 225
12.6.2 Combined Effect of Ozone and Chemical Treatment ............................................ 226
12.6.2.1 Organic Acids ........................................................................................ 226
12.6.2.2 Chlorine ................................................................................................. 227
12.6.2.3 Hydrogen Peroxide ................................................................................ 228
12.6.2.4 Negative Air Ions (NAI) ........................................................................ 228
12.7 Constraints in Application of Ozone in Food Industry ....................................................... 228
12.8 Future Perspectives ............................................................................................................. 228
References ...................................................................................................................................... 229
12.1 INtrODUCtION
Ozone (triatomic oxygen molecule) is formed by the addition of a free radical of oxygen to molec-
ular oxygen photo-chemically in the stratosphere of the earth’s atmosphere approximately 20–30 km
above the surface. It acts as a shield for living organisms on the earth from harmful ultraviolet rays
213
Sanitization
Surface Disinfection
sterilization water
Applications of
ozone
Surface
Decontamina- removal of
tion pests
Sterilization
of food
packaging
material
Figure 12.1 Application areas of ozone in food processing.
of solar radiation. Ozone can be used in food processing in several ways (Figure 12.1). Ozone is a
well-known and powerful broad-spectrum antimicrobial agent and is active against bacteria, fungi,
viruses, protozoa, and spores of bacteria and fungi; the microbicidal effect of ozone is attributed
to its higher oxidation potential (Foegeding, 1985; Restaino et al., 1995; Khadre et al., 2001). The
inactivation of bacterial cells by ozone is achieved by the gradual oxidation of vital cellular com-
ponents. First-stage oxidation takes place at the surface leading to the degradation of unsaturated
lipids of the cell envelope; in the next stage, the cell membrane barrier is destroyed causing the dis-
ruption of the cell with leakage of cellular contents, which leads to the bacterial cells’ lysis. Studies
by Komanapalli and Lau (1996) confirmed that ozone penetrates inside the bacterium and oxidizes
certain essential components such as enzymes, proteins, and nucleic acids required for its survival.
Application of ozone as a disinfectant in water treatment started long back, and its use in food
industry started in 2001 after the FDA gave the status of “generally recognized as safe” (GRAS)
to the ozone as a direct additive in food. Ozone is being used for several other purposes such as for
the preservation of meat, poultry (Sheldon and Brown, 1986), shrimp (Chen et al., 1992), and fruits
and vegetables (Badiani et al., 1996; Achen and Yousef, 1999), and also to oxidize toxic organic
compounds, to reduce the odour, surface cleaning in food industries, and to decrease the biological
oxygen demand in the environment. The use of ozone minimizes inorganic waste because the mol-
ecules decompose spontaneously to oxygen (Adler and Hill, 1950).
Food products are commonly treated with gaseous and aqueous forms of ozone. The form of ozone
treatment is determined by the types of food products being processed (Cullen et al., 2010). Various
researcher have advocated use of water containing ozone for higher efficacy (Kim et al., 1999; Qiang
et al., 2005; Zorlugenc et al., 2008); however, another group of researchers reported that gaseous ozone
is more efficient leading to higher microbial log reductions (Singh et al., 2002). These variable results
may be because of differences in commodities and treatment conditions used by researchers.
Ozone application at the post-harvest stage of fruit and vegetables is for inactivation of
pathogenic and spoilage-causing microorganisms, and reduction of pesticide and chemical resi-
dues. Whole fruits like apples, blackberries, blueberries, cantaloupe, cantaloupe melon, peaches,
oranges, etc. were studied for the action of ozone against different microorganisms in last decade.
Apples (McLoughlin, 2000) and sliced salad mixes (Strickland et al., 2010) are being treated indus-
trially with water containing ozone to increase shelf life. Gaseous ozone was also successfully
applied for microbial load reduction in various vegetables such as tomatoes, asparagus, broccoli,
cucumber, green beans, and lettuce. Rice (2005) reported that the ozone treatment resulted in the
prevention of fungal disease of onions and potatoes. Ozone was also used for surface discolouration
iMProVinG tHe eFFiCACy oF oZone treAtMent in FooD PreserVAtion 215
of broccoli florets (Lewis et al., 1996), spinach leaves (Sakaki et al., 1983), and peaches (Badiani
et al., 1996). Liew and Prange (1994) also reported that ozone-treated carrots were lighter in colour
as compared to the non-treated carrots. However, it caused the destruction of chlorophyll in fresh
romaine lettuce (Bermudez and Barbosa, 2013).
Use of ozone as a direct additive to the fruit juices for preservation was the topic of interest for
many research groups throughout the world as ozone processing has potential to become one of the
least expensive preservation techniques among non-thermal technologies. Antimicrobial efficacy of
gaseous ozone against bacterial species such as S. cerevisiae, E. coli, and Salmonella in orange and
apple juice was studied during the last decade (Williams et al., 2005; Patil et al., 2010a, 2010b; Choi
et al., 2012). The action of ozone in a liquid depends on its solubility in liquid and follows Hendry’s
law, which states that the partial pressure of the gas in a liquid (solubility) is directly proportional to
the partial pressure of the gas.
12.2 OZONE GENEratION MEthODS
Artificially, ozone can be produced by different methods such as corona discharge, cold-plasma,
and ultraviolet. In addition, ozone can also be produced by chemical, thermal, chemo-nuclear, and
electrolytic methods (Kim et al., 1999). The generation of ozone is an endothermic reaction, and the
application of heat can quickly degrade it. It is a thermally unstable molecule, and its decomposition
is exothermic.
( ∆H•at1 atm,+ 284.5 KJ / mol )

3O2 ←
→ 2O3 (12.1)
It has a blue colour when generated using dried air at room temperature but colourless when
generated from high-purity oxygen. Molecular ozone in gaseous as well as in water-soluble form
decomposes back to the oxygen. The pungent odour of ozone was first described by Van Marum in
the year 1780 (Evans, 1972) and later, in the year 1840, Schönbein named it as “ozone,” based on
the Greek word “ozein” for “smell” (Rice and Bollyky, 1981; Kogelschatz, 1988). The thermal gas-
phase decomposition was given by Oyama (2000) as:
O3 + M → O 2 + O + M (12.2)
O3+ O → 2O2 (12.3)
12.2.1 Corona Discharge Method
This method is based on passing a dried, oil-free, and dust-free oxygen-containing gas through a
high-energy electrical field separated by a dielectric material, which is usually a glass (Figure 12.2).
As oxygen molecules pass through the medium, they are made to split apart, forming free atomic
oxygen radicals that can combine with divalent oxygen molecules to produce molecular ozone.
The ozone/gas mixture generated from the Ozonator normally contains 1%–3% ozone when dry
air is used as the feed gas or 3%–6% ozone when highly pure oxygen is used as the feed gas
(Muthukumarappan et al., 2000; Cullen et al., 2010). Generation of ozone is represented as
O2 + e − → 2O (12.4)
2O + 2O2+ → 2O3 (12.5)

Figure 12.2 Corona discharge method of ozone generation.
12.2.2 photochemical Ozone Generator
UV-based ozone is generated using low-pressure mercury lamp. The mercury lamp emits two
high-efficiency resonance lines with wavelengths of 185 and 254 nm (Voronov, 2008). The 185 nm
wavelength is responsible for ozone production, whereas 254 nm wavelength is effective against
DNA, thereby disrupting the growth of an organism. The lamps are made up of quartz through which
feed gas (usually ambient air) passes through the high-energy irradiance lamp. Photo-disassociation
splits the oxygen molecules (O2) into unstable oxygen radical atoms (O1), which being unstable react
with surrounding oxygen molecules to form ozone (O3). The electrochemical method generates
ozone between anode and cathode by dipping in a solution containing water and a solution of highly
electronegative anions (Muthukumarappan et al., 2008, 2009; Cullen et al., 2010).
Ozone mixture with oxygen is relatively more stable in the gaseous form than the aqueous soluble
state. This can be judged by the longer half-life of ozone in gaseous form than the ozone dissolved
in water, which quickly degrades to oxygen. Decomposition of ozone in pure water occurs through a
complex radical chain mechanism (Wojtowicz, 1996), and the sequence is initiated by hydroxyl radical.
O3 + OH − → HO2 + O2 − (12.6)
Unstable nature and reactivity of the ozone are due to its structure. It contains three oxygen
atoms bound by rearrangement of the three sp2 hybrid orbitals, forming a triangle with an oxygen
nucleus at its centre, with an angle of 116° 49′ (Beltran, 2004). Because of this structure ozone is
a more potent oxidizing agent than molecular oxygen and hydrogen peroxide. Ozone has a high
oxidizing potential of 2.07 V compared to chlorine (1.36 V) and oxygen (1.23 V) (Pehkonen, 2001),
and it reacts with almost all substances at room temperature.
The action of the ozone on microbes is also dependent on the presence of organic matter in the
liquid foods (Guzel-Seydim et al., 2004). Many researchers discussed factors affecting the action
of ozone on liquid food and these were categorized as extrinsic factors (flow rate of the ozone gas,
the concentration of ozone, and temperature of the subjecting liquid) and intrinsic factors (ozone
consuming compounds, solid content, and pH of the subjecting liquid).
12.3 EXtrINSIC FaCtOrS
12.3.1 Concentration
Higher ozone concentration resulted in faster inactivation, shorter treatment time, and
smaller D values (Steenstrup and Floros, 2004). An ozone concentration of 0.90 mg/L reduced
the E. coli populations of approximately 1 × 108 cells/mL by 4 logs in 6 min, and the use
of higher ozone concentration (4.72 mg/L) resulted in approximately 5-log reduction in 4 min
(Patil et al., 2009a) indicating that the concentration governs the efficacy of ozone for microbial
inactivation. However, higher ozone concentration may cause oxidation of health-promoting
compounds.
12.3.2 temperature
The solubility of ozone in water is 13 times more than that of oxygen at 0°C–30°C, and it is more
soluble in colder water (Rice, 1986; Bablon et al., 1991). The increase in temperature renders ozone
less soluble and less stable but the decomposition rate increases (Rice et al., 1981). The ability of
ozone to inactivate bacteria decreases with decreasing temperature (Vaughn et al., 1987; Hunt and
Marinas, 1997), and increasing temperature (7°C–22°C) had the strongest influence on the inactiva-
tion rate of Bacillus subtilis spores (Dow et al., 2006). The simultaneous contribution of solubility/
stability and reactivity to ozone efficacy can vary with experimental conditions, making it difficult to
predict the influence of temperature on a particular application (Pascual et al., 2007). In general, at a
lower temperature, the ozone is more stable in aqueous soluble form, and as the temperature increases
the decomposition rate increases (Sease, 1975). Contrastingly, it was also reported that as the tempera-
ture increases the antimicrobial action of the ozone also increases, which suggests that the stability of
the ozone is compensated with the reactivity.
12.4 INtrINSIC FaCtOrS
12.4.1 ph
pH plays a significant role in the solubility as well as in the rate of decomposition of the
ozone. Kuscu and Pazir (2004) indicated that decrease in pH increases ozone efficacy, which
may be due to the change in ozone decomposition rate substantially with a decrease in pH. It
was reported that the kinetics of inactivation of E. coli was much faster in the acidic medium
than in basic medium. The inactivation of E. coli (108 CFU/mL) at a flow rate of 2 L/min with
an ozone concentration of 0.90 mg/L at 25°C± 2°C resulted in a higher rate constant at a pH
value of 4.93 than at pH 9.16 (Zuma et al., 2009). The dependency of ozone solubility on pH
was recently studied by Egorova et al. (2015), and they reported the highest solubility of ozone
in the range of 2.5–6.0 pH.
12.4.2 Organic Matter Content
Food components are reported to interfere with the antimicrobial properties of ozone.
Efficacy of ozone is demonstrated more readily when targeted microorganisms are suspended
and treated in pure water or simple buffers (with low ozone demand) than in complex food
systems in which it is difficult to predict how ozone reacts in the presence of organic matter.
Presence of organic substances with high ozone demand in a medium may compete with micro-
organisms for ozone. The presence of a high level of organic solid content provides protection to
microorganisms (Seydim et al., 2004). Patil et al. (2009a, 2009b) demonstrated that higher inac-
tivation rate of E. coli was obtained in model orange juice as compared to the unfiltered cloudy
juice, probably due to the presence of other constituents such as fibre, ascorbic acid, and sugars
in the cloudy juice, which acted as barriers to inactivation of the microorganisms (O’Donnell
et al., 2012).
12.5 OZONE aS a prESErVatIVE
Preservation of whole fruits and vegetables, as well as juices by application of ozone, was studied
by several researchers. Washing or dipping apples in ozone (concentration of 25 mg/L) resulted in
decreased E. coli count by 3.7 or 2.6 log CFU/g, respectively (Achen and Yousef, 2001). Rodgers et al.
(2004) demonstrated that washing apples for 5 min with 3 ppm ozonated water reduced pathogens
(E. coli and Listeria monocytogenes) to undetectable levels, which remained below the detection
limit during the nine days of storage at 4°C. Patil et al. (2010a, 2010b, 2010c) demonstrated that ozone
treatment could be used as an effective process for reducing bacteria in apple juice. Application of
ozone (0.048 mg ozone at a constant flow rate of 0.12 L/min) for 5 min resulted in a 5-log reduction
of E. coli inoculated to apple juice.
Barth et al. (1995) demonstrated that a continuous supply of 0.3 ppm gaseous ozone results in an
extension of the shelf life of blackberries due to its antifungal effect, and maintained the red colour
of the blackberries up to 12 days of storage at 2°C. Song et al. (2003) pointed out that ozone treat-
ment could be used to reduce decay of blueberry fruit, as the fruits subjected to 700 ppb of ozone for
four days followed by four weeks in controlled atmosphere storage resulted in a significant higher
marketability than that of the control. Barboni et al. (2010) demonstrated that ozone-treated kiwi
fruit stored for four to six months in industrial freezer chambers at 0°C led to decrease in firmness
and acidity; whereas reducing sugar, TSS, and pH was found to increase. Minas et al. (2010) demon-
strated that the ozone treatment blocked ethylene production and delayed ripening, and stimulated
antioxidant as well as antiradical activities of the fruit.
Silveira et al. (2010) demonstrated that melon pieces packed in polypropylene trays under a pas-
sive modified atmosphere (7.4 kPa O2 + 7.4 kPa CO2) and subjected to a combination of peracetic
acid with 0.4 mg/L of ozonated water (3 min) could be as effective as chlorine treatment. The ozone
treatment (0.3 ppm) resulted in delayed green and blue mould incidence on inoculated oranges for
about one week compared to ambient air, and the external mycelial growth and conidia development
of P. italicum and P. digitatum were prevented or reduced by the treatment (Palou et al., 2003, Renzo
et al., 2005). Further, the combination of these treatments with 0.25 ppm of ozone in the storage
atmosphere during the night decreased a load of spores in the storage room and inhibited the surface
growth of mould on packages, walls, and floors. Palou et al. (2003) showed that exposure of 0.72 ppm
ozone for 14 days was effective in reducing the ethylene level within the containers and retarding the
ripening process of the oranges, besides inhibiting the germination of spores of either P. digitatum or
P. italicum. Song et al. (2000) showed that ozone was also effective in reducing the airborne spore
concentration in the storage rooms while maintaining the firmness and reducing the sprouting and
rooting of onion bulbs. Fan et al. (2001) found that the exposure of the onion bulbs to 50 and 250 ppb
ozone during the day and night, respectively, reduced physiological weight loss and mould incidence
after four weeks of cold storage, and the shelf life of the product was extended up to an additional
two weeks. Tzortzakis et al. (2007, 2008) showed that tomatoes kept in ozone-enriched atmospheres
(0.05–5 mmol/mol) for 2–312 h inhibited spore production of B. cinerea, A. alternata, and C. coc-
codes by 94%–99%, 55%–80%, and 13%–74%, respectively. Elisabete et al. (2011, 2012) also reported
the use of ozone treatment in microbial load reduction (total mesophiles, and yeasts and moulds) and
quality retention (colour, firmness, pH, total anthocyanins, and ascorbic acid content) of strawberries.
Oner et al. (2011) indicated that in-package gaseous ozone treatment could be used for maintain-
ing quality and improving the shelf life of blanched potato strips, and continuous ozone treatment
was found to be effective in extending the shelf life of potato strips. Tiwari et al. (2008a, 2008b)
demonstrated that ozone treatment of orange juice showed no significant changes in pH, TSS, titrat-
able acidity, cloud value, or non-enzymatic browning. Further, the effect of gas flow rate, ozone
concentration, and treatment time on colour degradation on orange juice was investigated. Salient
results of various studies evaluating the effect of ozone on the physicochemical and nutritional
quality of food products are summarized in Table 12.1.
taBLE 12.1 Effect of Ozone on physico-chemical and Nutritional Quality parameters of Food products
Food product Ozone application Quality and Nutritional attributes references
orange juice Gaseous ozone Ascorbic acid (d), colour (s) Angelino et al. (2003)
orange juice Gaseous ozone Colour (d), non-enzymatic browning (nc), tiwari et al.
titratable acidity (nc), pH (nc), ascorbic acid (d) (2008a, 2008b)
Apple cider Gaseous ozone sediments (i), colour (s) Choi and nielsen (2005)
strawberry Gaseous ozone Colour (d), pH (nc), titratable acidity (nc), tiwari et al. (2009b)
juice ascorbic acid (d), anthocyanins (d)
blackberry juice Gaseous ozone Colour (d), anthocyanins (d) tiwari et al. (2009c)
tomato juice Gaseous ozone Colour (d), non-enzymatic browning (nc), tiwari et al. (2009a)
cloud value (nc), pH (nc), titratable acidity
(nc), ascorbic acid (d)
sugarcane Gaseous ozone Colour (i), non-enzymatic browning (d), Garud et al. (2017)
juice carotenes (nc), ascorbic acid (d)
Celery Aqueous ozone ascorbic acid (nc) Zhang et al. (2005)
blackberries Aqueous ozone Colour (nc), anthocyanin (nc) barth et al. (1995)
strawberry Gaseous ozone texture (s), Colour (s) Predmore et al. (2015)
Lettuce Gaseous ozone Colour (s) Predmore et al. (2015)
(s): significant difference; (i): increases; (d): decreases; and (nc): no change
Higher concentration of ozone used for preservation gives a fishy odour to the treated products,
besides oxidizing nutrients present in them, as the oxidation process by ozone is non-selective.
To overcome this problem researchers combined ozonation process with other antimicrobial treat-
ments to reduce the use level of ozone. The literature on the effectiveness of ozone in combination
with other treatments is described in following sections.
12.6 COMBINatION OF OZONE WIth OthEr hUrDLES

FOr ENhaNCING prESErVatION pOtENtIaL
The effectiveness of the ozone treatment can be enhanced by combining ozone with different
physical as well as chemical preservation techniques (Figure 12.3). Although only limited literature
is available on combined treatments using ozone, it confirms the concept of the synergistic effect of
ozone with other hurdles. These hurdles in combination with ozone can be categorized under two
major categories: physical hurdles and chemical hurdles. The following section discusses the com-
bination of ozone with physical as well chemical hurdles, and results of a few combination studies
on microbial quality are summarized in Table 12.2.
Figure 12.3 Possible combination treatments of ozone with physical and chemical hurdles.
220
table 12.2 Effect of Ozone in Combination with Other treatments on Microbial Quality of Juices
Ozone treatment Specification Combine hurdles hurdles treatment Specification results references
physical hurdles
Diffuser at bottom of plexi-glass ultrasound the ultrasonic system (40 KHz with inactivation of all bacterial strains burleson et al. (1975)
treatment column at a flow rate of 150 W) with transducer placed at with complete inactivation less
5 cubic ft per h the bottom of the Plexiglas treatment than one min
column.
ozone supply capable of generating the ultrasonic system 99% inactivation occurred after Al-Hashimi et al. (2015)
a concentration which accumulated (100 W/612 kHz), 16 min with pulsed 4 min treatment
to 1 ppm ultrasonic treatment on for 5 s and off
for 5 s
ozone (72–75 ppm) for up to 10 min ultrasound at amplitude level of 7.5 μm Combined ozone and sonication Patil (2010)
(flow rate of 0.12 L/min) reduced the population of E. coli
25922 in juice by 6.0 log in 4 min
Humidified ozone exposure ultraviolet ultraviolet (uV) treatment (200 and Combinations of ozone and uV Kim and Hung (2012)
(4000 ppm, 8 min) 280 nm) for 1, 5, and 10 min reduced the populations of E. coli
o157:H7 on the calyx of blueberries
by 3.35 log CFu/g
ozone (3, 6, and 24 g/m3) uV-C (253.7 nm) After 60 min treatments, total selma et al. (2008a)
mesophilic bacteria reductions
achieved by ozone–uV combinations
were 1.6–2.6 log CFu/mL higher than
by uV treatment alone
Aqueous ozone (4.5 ppm at 10°C) uV-C (254 nm, 1 kJ/m2) Combined application of ozone and Hassenberg et al.
uV caused highest reduction in yeast (2012)
loads of spears at the end of storage
Aqueous ozone (0.5 ppm) uV-C intensity of 1.19 mW/cm2 Combination of uV and ozonated Pang and Hung (2016)
water achieved an additional
1.8–2.7 log CFu/g reductions at
10–15 min of treatment compared
to either uV or ozonated water
treatment alone for the same
treatment times
(Continued)
table 12.2 (Continued) Effect of Ozone in Combination with Other treatments on Microbial Quality of Juices
ozone (up to 1 ppm in gas phase, PeF PeF (20 kV/cm) in bipolar wave form; L. leichmannii counts decreased by unal et al. (2001)
flow rate 11.4 mL/min) pulse frequency, 1,000 Htz; pulse 7.2 log10 CFu/mL by combination
duration time, 3 μs; number of chambers, of 1 ppm ozone and PeF
4; field strengths, 0–30 kV/cm; electrode
diameter, 0.23 cm; electrodes gap,
0.292 cm; and product flow rate, 1 mL/s.
the sample received approximately
12 pulses per chamber with total
treatment time of 145.6 μs
ozone in gas phase (10,000 ppm thermal treatment Whole melons were submerged in Combination achieved 3.8, 5.1 and selma et al. (2008b)
at 11°C for 30 min, 90%–95% 75°C water for 1 min 2.2 log reductions of mesophilic
relative humidity) bacteria, psychotropic bacteria,
and moulds, respectively
ozone (0.3 and 2.0 ppm) blanching at 50°C and 60°C for 3 min the log-reductions obtained did not Alexandre et al. (2011)
show synergistic effect with
combined treatments
ozone (0.9 g/h at gas flow rate temperature (50°C) up to 240 min Combination treatment reduced Williams et al. (2004)
of 2.4 L/min) treatment time to achieve 5-log
reductions of E. coli o157:H7 in apple
cider and orange juice
ozone (2.0–3.0 g/m3 for 1 min at temperatures (50°C and 55°C for 1 min) synergistic effect of combination for the sung et al. (2014)
flow rate of 3.0 L/min) inactivation of pathogens in apple juice
Chemical hurdles
ozone (2 ppm) organic acids Malic acid (2%) Combination reduced pathogen singla et al. (2011)
populations by 4.4 log in radish
and 4.8 log in moong bean sprouts
Aqueous ozone (3 ppm) Citric acid (1%) Combined treatment decrease yuk et al. (2006)
iMProVinG tHe eFFiCACy oF oZone treAtMent in FooD PreserVAtion
populations of E. coli o157:H7

compared with individual treatments
ozone (3 ppm) Citric acid (1%) Combined treatment resulted in yuk et al. (2007)
2.26- and 1.32-log count reductions
of E. coli o157:H7 and
L. monocytogenes, respectively
ozone (flow rate 1.2 gm/h for 10 min) Lactic acid (0.5%–1%) Higher reduction in natural microflora Garud et al. (2017)
of sugarcane juice by combined
treatment than individual treatments
(Continued)
221
222
table 12.2 (Continued) Effect of Ozone in Combination with Other treatments on Microbial Quality of Juices
ozone (2.5 ppm) Chlorine 100 ppm Combined treatment reduced the Garcia et al. (2003)
microbial load (by 1.4 log CFu/g) on
the lettuce better than individual
treatments
ozone (0.9 g/h, 2.4 L/min) Hydrogen peroxide 300 or 600 ppm reduction in Salmonella (5.9 log CFu/mL Williams et al. (2005)
in apple cider and 5.7 log CFu/mL in
orange juice) and E. coli o157:H7
(2.3 log CFu/mL in apple cider and
2.5 log CFu/mL in orange juice)
ozone (0.9 g/h, 2.4 L/min) Dimethyl 500 ppm More than 5-log reductions for Williams et al. (2005)
dicarbonate E. coli o157:H7 and Salmonella in
(DMDC) apple cider and orange juice
ozone (0.5 ppm) negative air ions 6.1 × 106 ions/cm3 Decrease in bacteria and yeasts count tuffi et al. (2012)
by 2.5 log10 CFu/g in tomatoes after
15 days storage at 12°C and 95% rH
12.6.1 Combined Effect of Ozone and physical treatment
12.6.1.1 Ultrasound
Different research groups have shown the synergistic action of ozone with ultrasound which
can be summarized into two main phenomena. The ultrasound causes reduction of interfacial dis-
tance (reduce gas-bubble film thickness) between the gas bubble and the liquid phase, which leads
to higher mass transfer across the phase and results in higher rate of microbial reduction. Another
mechanism proposes cell lysis caused by ultrasound decreasing the particle size of organic mat-
ter, which leads to more surface area to react for the ozone when used as a combined treatment.
Burleson et al. (1975) studied the effect of ozone and its combination with ultrasound (Figure 12.4)
and reported that the ozone treatment alone took longer time for the complete inactivation of micro-
organisms in both saline and secondary effluent than combined treatment. The combined treatment
involving ozonation and sonication was able to reduce the contact time for complete inactivation of
microorganisms. Al-Hashimi et al. (2015) also studied the effect of combined treatment (ultrasound
and ozone) for the bacterial disinfection of water on a large-scale application, and a synergistic
effect was seen with the combined treatment, which resulted in almost complete inactivation of
bacterial cells in 4 min. It was concluded that the combined ultrasound and ozone treatment is a
promising method for disinfection of water.
Figure 12.4 reduction in bacterial population by combination of ozone and sonication. (Modified from
burleson, G.r. et al., Appl. Microbiol., 29, 340–344, 1975.)
Use of ozone and ultrasound in combination was also studied for the degradation of the textile
dyes and pesticide residues by different research groups. Gultekin and Ince (2006) studied the
degradation of aryl-azo-naphthol dyes by ultrasound, ozone and their combinations. The results
indicated that the combined use of ultrasound and ozone improves the rate of bleaching and UV
absorption decay and enhances the mineralization of the dye. These effects were attributed to the
increased mass transfer of ozone in solution and its decomposition in the gas phase to yield hydroxyl
radicals and other oxidative species.
12.6.1.2 Ultraviolet (UV) Radiation
Ozone was found to have a synergistic effect when combined with UV light as it increases the
microbial reduction faster than the individual treatments. Kim and Hung (2012) inoculated blueber-
ries with five different E. coli cultures and treated with ozone, UV light, and the combination. It
was shown that the combined treatment gave better microbial reduction than individual treatments.
Selma et al. (2008a) used ozone and UV combined treatment for microbial reduction in the vegetable
washed water, and found that the combination was more suitable than the individual treatments for
microbial reduction in onion wash water (Figure 12.5). The synergy caused by a combination of
ozone and UV light may be due to the enhanced photocatalytic effect of UV radiation, which helps
in reducing the COD, and turbidity, and increases the efficacy of ozone. Pang and Hung (2016) com-
pared the efficacy of ozone in combination with UV light and slightly acidic electrolyzed water in
reducing the microbial count of E. coli O157:H7 cells on the Romaine and Iceberg Lettuce. Combine
treatment of ozone and UV light was found to give a higher reduction of microbial cells.
Diaz et al. (2001) elucidated the effect of various treatments on reducing the microbial contami-
nation of poultry processing chiller water. It was observed that ozone in combination with UV light
and oxygen was optimal treatment among the four treatments (O2/O3O2/UV, O2/O3/UV, and O2 as
the control). Combined treatment was found to give synergistic effect over individual treatment and
provided greater than 99.9% control of pathogenic microorganisms. Lin et al. (2012) studied the
combined ozone/UV/TiO2 treatment for the removal of three pesticides (cypermethrin, malathion,
dichlorovos) from fresh tea leaves. Removal of pesticides from tea leaves by ozone/UV/TiO2 combination
4
Log reduction
0
UV O3 UV+O3 UV O3 UV+O3 UV O3 UV+O3
Total mesophilic bacteria Total coliforms Yeasts
Figure 12.5 reduction of microflora by uV illumination, ozonation, and their combination in onion wash water.
(Modified from selma, M.V. et al., Food Microbiol., 25, 809–814, 2008a.)
was significantly improved, and the combined treatment resulted in 80% and 78% removal of cyper-
methrin and malathion, respectively.
12.6.1.3 Pulsed Electrical Field (PEF)
Pulsed electric field (PEF) is a non-thermal processing technique with potential applications in
the food industry. The combination of ozone and PEF showed a synergistic effect on inactivation of
microorganisms, probably on account of the fact that both ozone and PEF act on the cell membrane.
Ozone acts on the lipids of the microbial cell membrane and PEF causes electroporation, and there-
fore, the combined treatment was able to alter cell membrane rapidly and produce the bactericidal
effect. Unal et al. (2001) explored the combination of ozone (0.25–1.00 μg per mL of cell suspen-
sion) and PEF (10–30 kV/cm) treatments against selected food-borne bacteria (Lesmanialeichmannii
ATCC 4797, Escherichia coli O157:H7 ATCC 35150, and Listeria monocytogenes) and concluded
that exposure of L. leichmannii, E. coli, and L. monocytogenes to ozone followed by the PEF treat-
ment showed a synergistic bactericidal effect as it caused faster microbial inactivation as compared
to ozone or PEF alone. Ohshima et al. (1997) studied the effect of combination of ozone and PEF
for bactericidal effect against E. coli and showed that simultaneous application of PEF and ozone
showed a synergistic effect on inactivation of E. coli. The observations made by these researchers
suggest that use of ozone and PEF can provide synergy, and ozone treatment should be done either
before the PEF treatment or simultaneously with both treatments.
12.6.1.4 Thermal Treatment
Thermal treatment is a widely accepted and conventionally used treatment to attain microbial
safety. Although it is the most widely used technique, it leads to the undesirable sensorial and
nutritional changes, such as colour degradation, softening of tissues, vitamin losses, and devel-
opment of unpleasant cooked flavour. Alternatively, thermal and non-thermal treatments can be
used in combination to avoid loss of quality as well as to increase shelf life. Selma et al. (2008b)
showed the better effect of gaseous ozone and hot water in combination on the sensory and micro-
bial quality (growth evolution of aerobic mesophilic and psychrotrophic bacteria, coliforms, and
moulds) of cantaloupe melon. It was concluded that hot water, gaseous ozone, and their combination
were effective in reducing the total microbial population. The combination treatment was found
to be the most effective treatment to control microbial growth and maintain the sensory quality
of melons. Alexandre et al. (2011) studied the effectiveness of ozone (0.3 and 2.0 ppm) in aque-
ous solution and blanching for microbial inactivation in three microorganism/food combinations:
Listeria innocua/red bell peppers (artificially inoculated), total mesophiles/strawberries, and total
coliforms/watercress. It was observed that combining blanching and ozone did not generate syner-
gistic effects, and in some situations, microbial reductions were lower than the ones observed when
treatments were applied independently (Figure 12.6). Further, the study suggested gaseous ozone in
the combination of hot water can increase microbial reduction significantly, and aqueous ozone with
blanching did not show an increase in microbial reduction. Aqueous ozone at higher temperature
degrades faster than gaseous ozone, and this may not help in using aqueous ozone with thermal
treatment. Williams et al. (2004) indicated that mild heating (50°C) in combination with ozone
treatment reduced treatment time to achieve greater than 5-log reductions of E. coli O157:H7 in
apple cider and orange juice. Ozone in combination with mild heat (50°C) was shown as an alter-
native to thermal pasteurization. Similarly, Sung et al. (2014) investigated the combined effect of
ozone and thermal treatment for the inactivation of E. coli O157:H7, Salmonella typhimurium, and
Listeria monocytogenesin apple juice, and found that the combination treatment was more effective
as compared to individual treatments in the inactivation of foodborne pathogens while maintaining
acceptable apple juice quality.
5
Log reduction
0
B60-O3 O3-B60 B55-O3 O3-B55 B55-O3 O3-B55
L. innocua /Bell pepper Total Total coliforms/watercress
mesophiles/Strawberries
Figure 12.6 effect of combining blanching (b at 55°C or 60°C) with ozonated water washings (0.3 ppm) (o3)
on load log-reductions in various food microorganism combinations. (Modified from Alexandre,
e.M.C., J. Food Eng., 105, 277–282, 2011.)
12.6.2 Combined Effect of Ozone and Chemical treatment
12.6.2.1 Organic Acids
The individual treatment of organic acids has antimicrobial action because of lowering the
pH of the system. Reduction in pH favours undissociated acid molecules that interfere with the
cellular metabolism of microorganism, and it reduces microbial activity as a result of pH change
in the microbial cell’s environment (Booth, 1985). Radish and moong bean sprouts were treated
with ozone, malic acid, and their combinations (Singla et al., 2011), and the combined treatment
significantly reduced pathogen populations in both the sprouts as compared to the individual treat-
ment. The combination of ozone and malic acid was found effective to reduce bacterial population
on the food contact surface like PVC pipes, polyethylene bags, and plastic surfaces; it also reduced
the biofilm formation by S. typhimurium, indicating that the combined treatment can be used as an
effective disinfectant for food contact surfaces (Singla et al., 2014).
When organic acids were coupled with the ozone, the combination showed synergy in reduc-
ing the microbial count of gram-positive as well as gram-negative bacteria of lettuce and enoki
mushroom, which may be due to the reduction in pH, which leads to higher stability of ozone (Yuk
et al., 2006, 2007). The effect of individual organic acid (acetic acid, citric acid, and lactic acid) and
their combination with ozone against E. coli O157:H7 and L. monocytogenes on enoki mushrooms
(Figure 12.7) showed that lactic acid and citric acid in combination with ozone were the better com-
binations among all the organic acids as higher reduction in microbial cells were observed (Yuk
et al., 2006). Ascorbic acid did not show any significant difference in efficacy between stand-alone
treatment and in combination with the ozone.
A combination treatment of gaseous ozone (6 ppm), acetic acid (0.5%), and ozonated water
(26 mg/L) was found to be most effective for wheat grain disinfection and can be used to replace
the use of chlorinated water (Dhillon et al., 2010). Beltran et al. (2005) showed that the combined
treatment of ozonated water and peroxyacetic acid of fresh-cut potatoes stored under modified
atmosphere packaging retained initial texture and aroma, prevented browning, and controlled
Figure 12.7 effect of 5-min exposure to ozone (3 ppm) and organic acids (1%), and their combinations on the
populations of E. coli o157:H7 (a) and L. monocytogenes (b) on enoki mushrooms. oZ, ozone;
AA, acetic acid; CA, citric acid; LA, lactic acid. (Modified from yuk, H.G. et al., J. Food Sci., 71,
M83–M87, 2006.)
microbial growth besides maintaining sensory quality. However, the use of ozonated water alone
was not effective in reducing total microbial populations.
12.6.2.2 Chlorine
Chlorine has long been used for disinfection of water due to its high oxidizing potential,
which gives it antibacterial properties. Chlorine usage as bleaching/antimicrobial agent leads to
the generation of harmful chemical by-product. These adverse effects can be reduced by using
ozone in combination with chlorine. Singh et al. (2002) demonstrated that a log reduction of
1.48–1.97 log CFU/g in E. coli was obtained using aqueous chlorine dioxide (ClO2, 10.0 mg/L for
10 min), ozonated water (9.7 mg/L for 10 min), or thyme oil suspension (1.0 mL/L for 5 min) on
baby carrots. The change in the order of sequential washing treatments like thyme oil followed
by aqueous ClO2/ozonated water, or ozonated water/aqueous ClO2, resulted in 3.99- or 4.34-log
reduction, respectively. Garcia et al. (2003) demonstrated that combination of free available ozone
and chlorine was the best combination to wash/rinse fresh-cut iceberg lettuce and to reduce the
mesophiles and psychrotrophic microorganisms, which provided a shelf life of more than 25 days
besides improving the water quality in the processing line.
12.6.2.3 Hydrogen Peroxide
Hydrogen peroxide is a strong oxidizing/bleaching agent and disinfectant known for its antibacte-
rial properties. Sommer et al. (2004) studied the efficacy of ozone, hydrogen peroxide, and the combi-
nation of microorganism and viruses. Experiments were carried out at pilot scale on polluted ground
water, and the results showed that combination of ozone and hydrogen peroxide was more effective than
individual treatments. Williams et al. (2005) studied the effect of ozone in combination with hydrogen
peroxide (300 and 600 ppm) for inactivation of E. coli and Salmonella in apple cider and orange juice.
Combined treatment showed synergy at both concentrations of hydrogen peroxide. However, no signifi-
cant differences among the different concentrations of hydrogen peroxide were observed.
12.6.2.4 Negative Air Ions (NAI)
Negative air ions (NAI) have a physiological effect on living organisms (Krueger and Reed,
1976; Charry and Kavet, 1987; Reiter, 1993). Fan et al. (2002) studied the effect of combined treat-
ment of ozone with NAI on bacteria inoculated on different culture media. It showed that NAI alone
did not have any effect on the cell death but showed synergy when it was combined with ozone, and
death rate was found to be substantially greater than individual ozone treatment. Tanimura et al.
(1998) reported that ozone and NAI were more effective in combination than alone. Combination of
ozone and NIA was also found effective in reducing decays in the oranges when they were treated
with ozone and NAI daily for 10 min. Synergistic effect of ozone and NAI was reported in case of
table grapes also (Hildebrand et al., 2001). Tuffi et al. (2012) studied the effect of ozone, NAI, and
their combinations for the control of postharvest decay of cold-stored tomatoes, and reported that
NAI combined with ozone and electrostatic filters not only reduced microbial population but also
reduced the disease incidence during storage.
12.7 CONStraINtS IN appLICatION OF OZONE IN FOOD INDUStrY
A thorough literature search on applications of ozone for the preservation of fruit, vegetables, and
fruit juices shows that many researchers demonstrated the antimicrobial effect of ozone. However,
there are several constraints in using ozone in the food industry, which need to be resolved before
exploiting its commercial application. It has a characteristic pungent odour which may affect the
quality of the product. Further, inhalation of ozone is toxic and causes dryness of throat and mucous
membrane, headache, and irritation of nose even at a lower concentration. The most important con-
straint in the application of ozone in food products is its interaction with organic matter present in
foods, which hinders its efficacy and demands higher concentration for effective use.
12.8 FUtUrE pErSpECtIVES
Ozone has a strong microbicidal action against bacteria, fungi, parasites, and viruses, and it is
suitable for washing and sanitizing food items. Ozone has been used in the preservation of many
foods and food ingredients, disinfection, and several other applications. Readily available organic
constituents in food, however, compete with microorganisms for applied ozone and thus efficacy of
the treatment is minimized. The antimicrobial efficacy can be enhanced to a great extent by com-
bining ozone with other chemical or physical treatments. The food industry is interested in using
ozone to decontaminate processing water and decrease its chemical and biological oxygen demand.
Multi-functionality of ozone application, thus, makes it a promising agent for food uses.
rEFErENCES
Achen, M., and Yousef, A.E. (1999). Ozone Treatment of Apples to Reduce Eschericia Coli O157:H7
Contamination. Institute of Food Technologists Annual Meeting: Chicago, IL, p. 215.
Achen, M., and Yousef, A.E. (2001). Efficacy of ozone against Escherichia coli O157:H7 on apples. J. Food
Sci. 66:1380–1384.
Adler, M.G., and Hill, G.R. (1950). The kinetics and mechanism of hydroxide ion catalysed ozone decomposi-
tion in aqueous solution. J. Am. Chem. Soc. 72:1884–1886.
Alexandre, E.M.C., Santos-Pedro, D.M., Brandao, T.R.S., and Silva, C.L.M. (2011). Influence of aqueous
ozone, blanching and combined treatments on microbial load of red bell peppers, strawberries and
watercress. J. Food Eng. 105(2):277–282.
Al-Hashimi, A.M., Mason, T.J., and Joyce, E.M. (2015). Combined effect of ultrasound and ozone on bacteria
in water. Environ. Sci. Technol. 49(19):11697–11702. doi:10.1021/es5045437.
Angelino, P.D., Golden, A., and Mount, J.R. (2003). Effect of Ozone Treatment on Quality of Orange Juice.
Institute of Food Technologists Annual Meeting, Chicago, IL.
Bablon, G., Belamy, W.D., Billen, G., Bourbigot, M.M., Daniel, F.B., Erb, F., Gomella, C. et al. (1991). Practical
applications of ozone: Principles and case studies. In: Ozone in Water Treatment, pp. 133–316. Langlais,
B., Reckhow, D.A., and Brink, D.R., (Eds.), Lewis Publisher, Chelsea, MI.
Badiani, M., Fuhrer, J., Paolacci, A.R., and Giovannozzi, S.G. (1996). Deriving critical levels of ozone effects
on peach trees (Prunuspersica L. Batsch) grown in open-top chambers in Central Italy. Fresen. Environ.
Bull. 5:594–603.
Barboni, T., Cannac, M., and Chiaramonti, N. (2010). Effect of cold storage and ozone treatment on physico-
chemical parameters, soluble sugars and organic acids in Actinidiadeliciosa. Food Chem. 121:946–951.
Barth, M.M., Zhou, C., Mercier, J., and Payne, F.A. (1995). Ozone storage effects on anthocyanin content and
fungal growth in blackberries. J. Food Sci. 60:1286–1288.
Beltran, D., Selma, M.V., Tudela, J.A., and Gil, M.I. (2005). Effect of different sanitizers on microbial and
sensory quality of fresh-cut potato strips stored under modified atmosphere or vacuum packaging.
Postharvest Biol. Technol. 37:37–46.
Beltran, F.J. (2004). Ozone Reaction Kinetics for Water and Wastewater System. CRC Press, New York, 357p.
Bermudez, A.D., and Barbosa, G.V.C. (2013). Disinfection of selected vegetables under nonthermal treat-
ments: Chlorine, acid citric, ultraviolet light and ozone. Food Control 29:82–90.
Booth, I.R. (1985). Regulation of cytoplasmic pH in bacteria. Microbiol. Rev. 49:359–378.
Burleson, G.R., Murray, T.M., and Pollard, M. (1975). Inactivation of viruses and bacteria by ozone, with and
without sonication. Appl. Microbiol. 29(3):340–344.
Charry, J.M., and Kavet, R. (1987). Air Ions: Physical and Biological Aspects. CRC Press, Boca Raton, FL,
205p.
Chen, H.C., Huang, S.H., Moody, M.W., and Jiang, S.T. (1992). Bacteriocidal and mutagenic effects of ozone
on shrimp (Penaeusmondon) meat. J. Food Sci. 57:923–927.
Choi, L.H., and Nielsen, S.S. (2005). The effect of thermal and non-thermal processing methods on apple cider
quality and consumer acceptability. J. Food Quality 28:13–29.
Choi, M.R., Liu, Q., Lee, S.Y., Jin, J.H., Ryu, S.R., and Kang, D.H. (2012). Inactivation of Escherichia coli
O157:H7, Salmonella Typhimurium and Listeria monocytogenes in apple juice with gaseous ozone.
Food Microbiol. 32:191–195.
Cullen, P.J., Valdramidis, V.P., Tiwari, B.K., Patil, S., Bourke, P., and O’Donnell, C.P. (2010). Ozone process-
ing for food preservation: An overview on fruit juice treatments. Ozone Sci. Eng. 32:166–179.
Dhillon, B., Wiesenborn, D., Dhillon, H., and Wolf-Hall, C. (2010). Development and evaluation of a fluidized
bed system for wheat grain disinfection. J. Food Sci. 75(6): E372–E378.
Diaz, M.E., Law, S.E., and Frank, J.F. (2001). Control of pathogenic microorganisms and turbidity in poultry-
processing chiller water using UV-enhanced ozonation. Ozone Sci. Eng. 23(1):53–64.
Dow, S.M., Barbeau, B., Von Gunten, U., Chandrakanth, M., Amy, G., and Hernandez, M. (2006). The impact
of selected water quality parameters on the inactivation of Bacillus subtilis spores by mono-chloramine
and ozone. Water Res. 40:373–382.
Egorova, G.V., Voblikova, V.A., Sabitova, L.V., Tkachenko, I.S., Tkachenko, S.N., and Lunin, V.V. (2015).
Ozone solubility in water. Moscow Univ. Chem. Bull. 70(5):207–210.
Elisabete, M.C.A., Dora, M.S.P., Teresa, R.S., Brandao, C., and Silva, L.M. (2011).Influence of aqueous ozone,
blanching and combined treatments on microbial load of red bell peppers, strawberries and watercress.
J. Food Eng. 105:277–282.
Elisabete, M.C.A., Teresa, R.S., Brandao, C., and Silva, L.M. (2012). Efficacy of non-thermal technologies
and sanitizer solutions on microbial load reduction and quality retention of strawberries. J. Food Eng.
108:417–426.
Evans, F.L. (1972). Ozone in Water and Wastewater Treatment. Ann Arbor Science Publishers, Ann Arbor, MI,
195p.
Fan, L.H., Song, J., Hildebrand, P.D., and Forney, C.F. (2001). Corona discharge reduces mold on commer-
cially stored onions. Acta Hortic. 553:427–428.
Fan, L., Song, J., Hildebrand, P.D., and Forney, C.F. (2002). Interaction of ozone and negative air ions to con-
trol micro-organisms. J. Appl. Microbiol. 93:144–148.
Foegeding, P.M. (1985). Ozone inactivation of Bacillus and Clostridium spore populations and the importance
of the spore coat to resistance. Food Microbiol. 2:123–134.
Garcia, A., Mount, J.R., and Davidson, P.M. (2003). Ozone and chlorine treatment of minimally processed
lettuce. J. Food Sci. 68(9):2747–2751.
Garud, S.R., Priyanka, B.S., Rastogi, N.K., Prakash, M., and Negi, P.S. (2017). Effect of non-thermal hurdles
on sugarcane juice preservation. Ozone Sci. Eng. doi:10.1080/01919512.2017.1415802.
Gultekin, I., and Ince, N.H. (2006). Degradation of aryl-azo-naphthol dyes by ultrasound, ozone and their
combination: Effect of alpha-substituents. Ultrason. Sonochem. 3(3):208–214.
Guzel-Seydim, Z., Bever, P.I., and Greene, A.K. (2004). Efficacy of ozone to reduce bacterial populations in
the presence of food components. Food Microbiol. 21:475–479.
Hassenberg, K., Huyskens-Keil, S., and Herppich, W.B. (2012). Impact of postharvest UV-C and ozone treat-
ments on microbiological properties of white asparagus (Asparagus officinalis L.). J. Appl. Bot. Food
Qual. 85:174–181.
Hildebrand, P.D., Song, J., Forney, C.F., Renderos, W.E., and Ryan, D.A.J. (2001).Effects of corona dis-
charge on decay of fresh fruits and vegetables. Proceedings of the Fourth International Conference on
Postharvest Science. Acta Hortic. 553:425–426.
Hunt, N.K., and Marinas, B.J. (1997). Kinetics of Escherichia coli inactivation with ozone. Water Res.
31:1355–1362.
Khadre, M.A., Yousef, A.E., and Kim, J. (2001). Microbiological aspects of ozone applications in food: A
review. J. Food Sci. 6:1242–1252.
Kim, C., and Hung, Y.C. (2012).Inactivation of E.coli O157:H7 on blueberries by electrolyzed water, ultravio-
let light, and ozone. J. Food Sci. 77(4): M206–M211.
Kim, J., Yousef, A., and Dave, S. (1999). Application of ozone for enhancing the microbiological safety and
quality of foods: A review. J. Food Prot. 62:1071–1087.
Kogelschatz, U. (1988). Advanced ozone generation. In: Process Technologies for Water Treatment,
pp. 87–120. Stucki, S., (Ed.), Plenum Publishers, New York.
Komanapalli, I.R., and Lau, B.H.S. (1996). Ozone-induced damage of Escherichia coli K-12. Appl. Microbiol.
Biotechnol. 46(5–6):610–614.
Krueger, A.P., and Reed, E.J. (1976). Biological impact of small air ions. Science. 193:1209–1213.
Kuscu, A., and Pazir, F. (2004). Ozone applications in food industry. Gida. 29 (2):123–129.
Lewis, L., Zhuang, H., Payne, F.A., and Barth, M.M. (1996). Betacarotene Content and Color Assessment in
Ozone Treated Broccoli Florets During Modified Atmosphere Packaging. Institute of Food Technologists
Annual Meeting, Chicago, IL.
Liew, C.L., and Prange, R.K. (1994). Effect of ozone and storage temperature on postharvest diseases and
physiology of carrots (Daucuscarota L.). J. Am. Soc. Hort. Sci. 119:563–567.
Lin, L., Xie, M., Liang, Y., He, Y., Chan, Y.S., and Luan, T. (2012). Degradation of cypermethrin, malathion
and dichlorvos in water and on tea leaves with ozone/UV/TiO2 treatment. Food Control 28:374–379.
McLoughlin, G. (2000). Apple processor switches from chlorine to ozone treatment. Water Technol. 23(2):53–54.
Minas, I.S., Karaoglanidis, G.S., Manganaris, G.A., and Vasilakakis, M. (2010). Effect of ozone applica-
tion during cold storage of kiwifruit on the development of stem-end rot caused by Botrytis cinerea.
Postharvest Biol. Technol. 58:203–210.
Muthukumarappan, K., Halaweish, F., and Naidu, A.S. (2000). Ozone. In: Natural Food Anti-Microbial
Systems, pp. 783–800. Naidu, A.S., (Eds.), CRC Press, Boca Raton, FL.
Muthukumarappan, K., O’Donnell, C.P., and Cullen, P.J. (2008). Ozone utilization. In: Encyclopedia of
Agricultural, Food, and Biological Engineering, pp. 1–4. Heldman, D.R., and Moraru, C.L., (Eds.),
CRC Press, Boca Raton, FL.
Muthukumarappan, K., O’Donnell, C.P., and Cullen, P.J. (2009). Ozone treatment of food materials. In: Food
Processing Operations Modelling Design and Analysis, pp. 266–267. Jun, S., and Irudayaraj, J.M.,
(Eds.), CRC Press, Boca Raton, FL.
O’Donnell, C., Tiwari, B.K., Cullen, P.J., and Rice, R. (2012). Ozone in Food Processing. Wiley Blackwell,
West Sussex, UK, 286p.
Ohshima, T., Sato, K., Terauchi, H., and Sato, M. (1997). Physical and chemical modifications of high-voltage
pulse sterilization. J. Electrostatics. 42:159–166.
Oner, M.E., and Walker, P.N. (2011). Effect of processing and packaging conditions on quality of refrigerated
potato strips. J. Food Sci. 76:35–40.
Oyama, T.S. (2000). Chemical and catalytic properties of ozone. Catalysis Rev. 42(3):279–322.
Palou, L., Smilanick, J.L., Crisosto, C.H., Mansour, M., and Plaza, P. (2003). Ozone gas penetration and
control of sporulation of Penicilliumdigitatum and Penicilliumitalicum within commercial packages of
oranges during cold storage. Crop Prot. 22:1131–1134.
Pang, Y.H., and Hung, Y.C. (2016).Efficacy of slightly acidic electrolyzed water and UV-ozonated water com-
bination for inactivating Escherichia Coli O157:H7 on romaine and iceberg lettuce during spray wash-
ing process. J. Food Sci. 81 (7): M1743–M1748.
Pascual, A., Llorca, I., and Canut, A. (2007). Use of ozone in food industries for reducing the environmental
impact of cleaning and disinfection activities. Trends Food Sci. Technol. 18: S29–S35.
Patil, S., Bourke, P., Frias, J.M., Tiwari, B.K., and Cullen, P.J. (2009a). Inactivation of Escherichia coli in
orange juice using ozone. Innov Food Sci. Emerg. Technol. 10(4):551–557.
Patil, S., Cullen, P.J., Kelly, B., Frias, J.M., and Bourke, B.P. (2009b). Extrinsic control parameters for
ozone inactivation of Escherichia coli using a bubble column. J. Appl. Microbiol. 107(3):830–837.
doi:10.1111/j.1365-2672.2009.04255.x.
Patil, S., Torres, B., Tiwari, B.K., Wijngaard, H.H., Bourke, P., Cullen, P.J., O’Donnell, C.P., and Valdramidis,
V.P. (2010b). Safety and quality assessment during the ozonation of cloudy apple juice. J. Food Sci.
75(7): M437–M443.
Patil, S., Valdramidis, V.P., Cullen, P.J., Frias, J., and Bourke, P. (2010a). Inactivation of Escherichia coli by
ozone treatment of apple juice at different pH levels. Food Microbiol. 27:835–840.
Pehkonen, A. (2001). The effect of dissolved ozone on the corrosion behaviour of some stainless steels
(dissertation). Helsinki University of Technology, Department of Materials Science and Rock
Engineering.
Predmore, A., Sanglay, G., Li, J., and Lee, K. (2015). Control of human norovirus surrogates in fresh foods by
gaseous ozone and a proposed mechanism of inactivation. Food Microbiol. 50:118–125.
Qiang, Z.M., Demirkol, O., Ercal, N., and Adams, C. (2005). Impact of food disinfection on beneficial biothiol
contents in vegetables. J. Agric. Food Chem. 53(25):9830–9840.
Reiter, R. (1993). Possible biological effects of electric and magnetic parameters in the environment.
Experientia. 49:769–774.
Renzo, G.C.D., Altieri, G., Derchia, L., Lanza, G., and Strano, M.C. (2005). Effects of gaseous ozone exposure
on cold stored orange fruit. Acta Hortic. 682:1605–1610.
Restaino, L., Frampton, E.W., Hemphill, J.B., and Palnikar, P. (1995). Efficacy of ozonated water against vari-
ous food-related microorganisms. Appl. Environ. Microbiol. 61 (9):3471–3475.
Rice, R.G. (1986). Application of ozone in water and waste water treatment. In: Rice, R.G., Browning, M.J.
(Eds.), Analytical Aspects of Ozone Treatment of Water and Waste Water, pp. 7–26. Syracuse, The
Institute, New York.
Rice, R.G. (2005). IOA-PAG user success reports: Commercial applications of ozone in agri-foods. In:
International Ozone Association Pan American Group Conference, Georgia, 9–12 October 2005.
Rice, R.G., and Bollyky, L.J. (1981). Fundamental aspects of ozone technology, in Rice, R.G. (Ed.) Ozone
Treatment of Water for Cooling Applications, pp. 1–19. Vienna, VA: The International Ozone
Association.
Rodgers, S.L., Cash, J.N., Siddiq, M., and Ryser, E.T. (2004). A comparison of different chemical sanitizers for
inactivating Escherichia coli O157: H7 and Listeria monocytogenes in solution and on apples, lettuce,
strawberries, and cantaloupe. J. Food Prot. 67:721–731.
Sakaki, T., Kondo, N., and Sugahara, K. (1983). Destruction breakdown of photosynthetic pigments and lipids
in ozone-fumigated spinach leaves with ozone-fumigation: Role of active oxygens. Physiol. Plant. 59
(1):28–34.
Sease, W.S. (1975). Ozone mass transfer and contact system. In: Rice R. G., Pichet, P., Vincent, M. A., (Eds.).
Proceedings of the Second International Symposium on Ozone Technology. International Ozone
Association, pp. 1–14, May 11–14, Stanford CT: International Ozone Association. Montreal, Canada.
Stanford, CT: International Ozone Association.
Selma, M.V., Allende, A., Lopez, G.F., Conesa, M.A., and Gil, M.I. (2008a). Disinfection potential of ozone,
ultraviolet-C and their combination in wash water for the fresh cut vegetable industry. Food Microbiol.
25:809–814.
Selma, M.V., Ibanez, A.M., Allende, A., Cantwell, M., and Suslow, T. (2008b). Effect of gaseous ozone and
hot water on microbial and sensory quality of cantaloupe and potential transference of Escherichia coli
O157:H7 during cutting. Food Microbiol. 25:162–168.
Seydim, G.Z.B., Greene, A.K., and Seydim, A.C. (2004). Use of ozone in the food industry. LWT Food Sci.
Technol. 37:453–460.
Sheldon, B.W., and Brown, A.L. (1986). Efficacy of ozone as a disinfectant for poultry carcasses and chill
water. J. Food Sci. 5:305–309.
Silveira, A.C., Aguayo, E., and Artes, F. (2010). Emerging sanitizers and clean room packaging for improving
the microbial quality of fresh-cut “Galia” melon. Food Control. 21:863–871.
Singh, N., Singh, R.K., Bhunia, A.K., and Stroshine, R.L. (2002). Efficacy of chlorine dioxide, ozone, and
thyme essential oil or a sequential washing in killing Escherichia coli O157: H7 on lettuce and baby
carrots. LWTFood Sci. Technol. 35(8):720–729.
Singla, R., Ganguli, A., and Ghosh, M. (2011). An effective combined treatment using malic acid and ozone
inhibits Shigella spp. on sprouts. Food Control. 22:1032–1039.
Singla, R., Goel, H., and Ganguli, A. (2014). Novel synergistic approach to exploit the bactericidal efficacy
of commercial disinfectants on the biofilms of Salmonellaentericaserovar Typhimurium. J. Biosci.
Bioeng. 118 (1):34–40. doi:10.1016/j.jbiosc.2013.12.025.
Sommer, R., Pribil, W., Pfleger, S., Haider, T., Werderitsch, M., and Gehringer, P. (2004). Microbicidal effi-
cacy of an advanced oxidation process using ozone/hydrogen peroxide in water treatment. Water Res.
Technol. 50 (1):159–164.
Song, J., Fan, L., Forney, C.F., Jordan, M.A., Hildebrand, P.D., Kalt, W., and Ryan, D.A.J. (2003). Effect of
ozone treatment and controlled atmosphere storage on quality and phytochemicals in highbush blueber-
ries. Acta Hort. 600:417–423.
Song, J., Fan, L., Hildebrand, P.D., and Forney, C.F. (2000). Biological effects of corona discharge on onions
in a commercial storage facility. Hort. Technol. 10:608–610.
Steenstrup, L.D., and Floros, J.D. (2004). Inactivation of Escherichia coli O157: H7 in apple cider by ozone at
various temperatures and concentrations. J. Food Process. Preserv. 28:103–116.
Strickland, W., Sopher, C.D., Rice, R.G., and Battles, G.T. (2010). Six years of ozone processing of fresh cut
salad mixes. Ozone Sci. Eng. 32(1):66–70.
Sung, H., Song, W., Kim, K., Ryu, S., and Kang, D. (2014). Combination effect of ozone and heat treatments
for the inactivation of Escherichia coli O157: H7, Salmonella Typhimurium, and Listeria monocyto-
genes in apple juice. Int. J. Food Microbiol. 171:147–153.
Tanimura, Y., Hirotsuji, J., and Tanaka, K. (1998). Food preservation technique using a mixed gas containing
negative ions and ozone. Shokuhin Kogyo (Food Industry). 41 (10):71–77.
Tiwari, B.K., Muthukumarappan, K., O’Donnell, C.P., and Cullen, C.P. (2008a). Kinetics of freshly squeezed
orange juice quality changes during ozone processing. J. Agric. Food Chem. 56:6416–6422.
Tiwari, B.K., Muthukumarappan, K., O’Donnell, C.P., and Cullen, P.J. (2008b). Modelling colour degrada-
tion of orange juice by ozone treatment using response surface methodology. J. Food Eng. 88:553–560.
Tiwari, B.K., O’Donnell, C.P., Brunton, N.P., and Cullen, P.J. (2009a). Degradation kinetics of tomato juice
quality parameters by ozonation. Int. J. Food Sci. Technol. 44(6):1199–1205.
Tiwari, B.K., O’Donnell, C.P., Muthukumarappan, K., and Cullen, P.J. (2009c). Anthocyanin and colour deg-
radation in ozone treated blackberry juice. Innov. Food Sci. Emerg. Technol. 10:70–75.
Tiwari, B.K., O’Donnell, C.P., Patras, A., Brunton, N.P., and Cullen, P.J. (2009b). Effect of ozone processing
on anthocyanins and ascorbic acid degradation of strawberry juice. Food Chem. 113 (4):1119–1126.
Tuffi, R., Lovino, R., Canese, S., Cafiero, L.M., and Vitali, F. (2012). Effects of exposure to gaseous ozone and
negative air ions on control of epiphytic flora and the development of Botrytis cinerea and Penicillium
expansum during cold storage of strawberries and tomatoes. Italian J. Food Sci. 24:102–114.
Tzortzakis, N., Borland, A., and Singletoni, B.J. (2007). Impact of atmospheric ozone-enrichment on quality-
related attributes of tomato fruit. Postharvest Biol. Technol. 45:317–325.
Tzortzakis, N., Singleton, I., and Barnes, J. (2008). Impact of low-level atmospheric ozone-enrichment on
black spot and anthracnose rot of tomato fruit. Postharvest Biol. Technol. 47:1–9.
Unal, R., Kim, J.G., and Yousef, A.E. (2001). Inactivation of Escherichia coli O157:H7, Listeria monocytogenes,
and Lactobacillus leichmannii by combinations of ozone and pulsed electric field. J. Food Prot.
6:777–782.
Vaughn, J.M., Chem, Y.S., Lindburg, K., and Morales, D. (1987). Inactivation of human and simian rotavi-
ruses by ozone. Appl. Env. Microbiol. 53:2218–2221.
Voronov, A. (2008). New generation of low pressure mercury lamps for producing ozone. Ozone Sci. Eng.
30:395–397.
Williams, R.C., Sumner, S.S., and Golden, D.A. (2004). Survival of Escherichia coli O157:H7 and Salmonella
in apple cider and orange juice as affected by ozone and treatment temperature. J. Food Prot.
67(11):2381–2386.
Williams, R.C., Sumner, S.S., and Golden, D.A. (2005). Inactivation of Escherichia coli O157: H7 and
Salmonella in apple cider and orange juice treated with combinations of ozone, dimethyl dicarbonate,
and hydrogen peroxide. J. Food Sci. 70(4): M197–M201.
Wojtowicz, J.A. (1996). Ozone. In: Encyclopedia of Chemical Technology, pp. 953–994. Othmer, K. (Eds.),
John Wiley & Sons, New York.
Yuk, H.G., Yoo, M.Y., Yoon, J.W., Marshall, D.L., and Oh, D.H. (2007). Effect of combined ozone and organic
acid treatment for control of Escherichia coli O157:H7 and Listeria monocytogenes on enoki mush-
room. Food Control 18:548–553.
Yuk, H.G., Yoo, M.Y., Yoon, J.W., Moon, K.D. Marshall, D.L., and Oh, D.H. (2006). Effect of combined ozone
and organic acid treatment for control of Escherichia coli O157:H7 and Listeria monocytogenes on
lettuce. J. Food Sci. 71(3): M83–M87.
Zhang, L.K., Lu, Z.X., Yu, Z.F., and Gao, X. (2005). Preservation of fresh-cut celery by treatment of ozonated
water. Food Control. 16(3):279–283.
Zorlugenc, B., Zorlugenc, F.K., Oztekin, S., and Evliya, I.B. (2008). The influence of gaseous ozone and
ozonated water on microbial flora and degradation of aflatoxin B (1) in dried figs. Food Chem. Toxicol.
46(12):3593–3597.
Zuma, F., Lin, J., and Jonnalagadda, S.B. (2009). Ozone-initiated disinfection kinetics of Escherichia coli in
water. J. Env. Sci. Health Part A. 44:48–56.
ChaptEr 13
high-pressure CO2 processing of Foods
Bindvi Arora, Alka Joshi, and Shruti Sethi
CONtENtS
13.1 Introduction ......................................................................................................................... 236

13.2 High-Pressure Carbon Dioxide ........................................................................................... 236
13.3 Mechanism of Action .......................................................................................................... 237
13.3.1 Displacement of Oxygen ....................................................................................... 237
13.3.2 Solubilization and Acidification ............................................................................ 237
13.3.3 Cell Rupture .......................................................................................................... 238
13.4 Advantages of the Technology ............................................................................................ 238
13.5 Effect on Microorganisms .................................................................................................. 238
13.5.1 Factors Affecting Microbial Inactivation .............................................................. 238
13.6 Enzyme Inactivation ........................................................................................................... 239
13.6.1 Mechanism of Action ............................................................................................ 239
13.6.1.1 Inhibitory Effects of Molecular CO2 .................................................... 239
13.6.1.2 The pH Lowering Effect.......................................................................240
13.6.1.3 Effects of Depressurization ..................................................................240
13.6.1.4 Possible Other Mechanisms .................................................................240
13.6.2 Factors Affecting Enzyme Inhibition....................................................................240
13.6.2.1 Effects of Pressure Levels .................................................................... 241
13.6.2.2 Effect of Temperature ........................................................................... 241
13.6.2.3 Effect of Treatment Time ..................................................................... 242
13.6.2.4 Effect of pH, Food Ingredients, and Processing Media ....................... 242
13.6.2.5 Effect of Microbubbled CO2 ................................................................. 242
13.6.2.6 Pressure Cycle ...................................................................................... 242
13.7 Application of HPCD in Beverage Industry ....................................................................... 242
13.8 Extraction of Nutraceuticals ...............................................................................................244
13.8.1 Effect of Temperature............................................................................................244
13.8.2 Effect of Pressure .................................................................................................. 245
13.8.3 Extraction of Carotenoids...................................................................................... 245
13.8.4 Extraction of Flavonoids ....................................................................................... 245
13.8.5 Extraction of Flavoring Compounds .....................................................................246
13.8.6 Encapsulation Using SC-CO2 ................................................................................ 247
235
13.9 Supercritical Fluid Extrusion ............................................................................................ 247

13.9.1 Bakery .................................................................................................................248
13.9.2 Extruded Snacks ................................................................................................. 249
13.9.3 Texturization Using SCFX.................................................................................. 249
13.9.4 Polymer Foaming ................................................................................................ 250
13.10 Effect on Food Quality Attributes .................................................................................... 250
13.11 Application of HPCD for Water Disinfection ................................................................... 251
13.12 Disinfection of Stored Grains ........................................................................................... 251
13.13 Limitations of the Technology .......................................................................................... 252
13.14 Regulatory Aspects ........................................................................................................... 253
References ...................................................................................................................................... 253
13.1 INtrODUCtION
Thermal processing is a widely used food preservation technique for pasteurization and ster-
ilization of foods. The treatment may be harmful for heat-sensitive food products and may also
impart undesirable organoleptic changes along with detrimental effects to the nutritional value of the
foods (Garcia-Gonzalez et al., 2007). Increasing consumer awareness for additive-free, shelf stable
food products with natural flavors, and nutrients has led the food industry to venture into nonther-
mal preservation technologies (Mertens and Knorr, 1992) such as high hydrostatic pressure (HHP),
high-pressure carbon dioxide (HPCD), pulsed electric field (PEF), ultrasonication, etc. Among these
non-thermal approaches, high-pressure carbon dioxide is being given a lot of focus lately (Damar
and Balaban, 2006). Compared to conventional heat pasteurization, HPCD avoids drawbacks such
as loss of flavor, denaturation of nutrients, and production of toxic compounds as well as changes in
physico-chemical and mechanical properties of the treated food. Fraser (1951) and Foster et al. (1962)
first reported about the disruption of bacterial cells by the rapid release of CO2 gas from a pressure
of about 34 atm to ambient pressure. After their experiments, Swift & Co. (Chicago, IL) obtained the
US patent in 1969 for sterilization of food with CO2 (Kauffman et al., 1969).
HPCD, also known as dense phase carbon dioxide (DPCD), is a non-thermal pasteurization/
sterilization technique that utilizes carbon dioxide (CO2) under high pressures (<50 MPa) to eliminate
spoilage and pathogenic microorganisms, inactivate enzymes, and protect the food from the detri-
mental effects of thermal treatment. Other possible applications of HPCD also include the improved
functionality of beverages, extraction of bioactives, extruded snacks, polymer development, water
disinfection, removal of storage pests, etc. The process involves use of gaseous CO2 under pressure,
liquid CO2, and supercritical CO2 (Balaban and Duong, 2014) to either solid or liquid foods to achieve
desirable changes. HPCD utilizes carbon dioxide (CO2), which is an inert, inexpensive, non-toxic gas
that has been given the status of a “generally recognized as safe” (GRAS) solvent (Ferrentino and
Spilimbergo, 2011). This chapter comprehensively details the role of the HPCD technique in various
food applications for microbial inactivation, enzyme inactivation, extraction of bioactives, and other
functions along with its regulatory concerns.
13.2 hIGh-prESSUrE CarBON DIOXIDE
Carbon dioxide has been used in the food industry for a long time as a constituent of gaseous
atmosphere in carbonated beverages and in modified atmosphere packed products. It is considered to
be inhibitory to microorganisms, with the effect being more pronounced when used under high pres-
sure (Haas et al., 1989). CO2 exists in all the three phases at its triple point of 517.77 kPa at −56.6°C
(Figure 13.1). Liquid CO2 is formed only at pressures above its critical pressure. The critical point for
CO2 is 7.38 × 103 kPa at 31.1°C, beyond which it can be used in supercritical applications. Its critical
HiGH-Pressure Co2 ProCessinG oF FooDs 237
Figure 13.1 Phase diagram of carbon dioxide.
temperature (31.1°C) is only slightly above room temperature, so detrimental effects of thermal
degradation are eliminated when the process is operated around the critical temperature. The lethal
effects of HPCD are found to be dependent on the pressure and temperature of operation (Lin et al.,
1993; Hong et al., 1997; Hong and Pyun, 1999) and water activity (aw) of the medium (Thakur et al.,
2013). Earlier, Haas et al. (1989) stated that a HPCD treatment would not be applicable to dry sub-
stances. In contrast to this, experiments conducted later by Dillow et al. (1999) concluded that E. coli
could be totally inactivated in the presence as well as in the absence of free water at 14 MPa at 34°C.
More recently, results of the study conducted by Calvo and Torres (2010) showed that microbial inac-
tivation during HPCD treatment was a strong function of the water activity.
13.3 MEChaNISM OF aCtION
Different theories have been put forth over the years regarding the mechanism of action of
HPCD, as discussed in the following sections.
13.3.1 Displacement of Oxygen
Carbon dioxide can completely displace oxygen present in foods, creating an anaerobic environ-
ment that interferes with key synthetic processes in the cell and proves detrimental to the contami-
nating microflora.
13.3.2 Solubilization and acidification
Carbon dioxide on reaction with water leads to formation of carbonic acid (H2CO3), which fur-
ther disassociates to produce bicarbonate ion (HCO3−) and hydrogen ion (H+), leading to reduction in
the pH of the medium. This inhibits microbial growth (Valley and Rettger, 1927; Daniels et al., 1985;
Hutkins and Nannen, 1993) and reduces the microbial resistance to inactivation because of increased
energy consumption to maintain pH homeostasis (Hong and Pyun, 1999). The low pH conditions
alter the enzymatic processes and cellular metabolism by causing an imbalance in the intracellular
electrolytes. In the supercritical state, CO2 has low viscosity and zero surface tension (McHugh and
Krukonis, 1993), so it can quickly penetrate the cell membrane of the microbial cells and cause
reduction in the intracellular pH leading to inhibition of enzymes and metabolic activities resulting
in microbial cell death (Hutkins and Nannen, 1993).
13.3.3 Cell rupture
Another theory envisages the effect of HPCD because of the increased internal pressure resulting
in cell rupture and the resultant release of vital constituents of the cell (Damar and Balaban, 2006).
13.4 aDVaNtaGES OF thE tEChNOLOGY
HPCD processing offers the following advantages:
1. Since CO2 is inexpensive and is readily available, it is easy to switch over to CO2-based pasteurization.
2. CO2 requires no special handling or ventilation.
3. CO2 can be easily removed by simple depressurization and degassing.
4. It leaves no toxic residues.
5. It is a good option for sterilization of heat-sensitive and/or porous biomaterials since operating tem-
peratures are not very high.
6. CO2 has an inhibitory effect on microorganisms and enzymes in the product.
7. Process time for extraction of compounds using super critical CO2 is reduced drastically in compari-
son to hydro-distillation.
8. Supercritical extraction achieves a good recovery of aromas and therefore yields an organoleptically
better tasting product.
Because of flexibility of operating conditions, CO2 has been proposed for use in biomaterial
applications such as preservation of foods, incorporation of bioactive ingredients into biodegradable
polymers (Cooper, 2001), and producing enzyme particles (Moshashaee et al., 2000).
13.5 EFFECt ON MICrOOrGaNISMS
Carbon dioxide gas has been shown to have bacteriostatic effect on microorganisms. Both
Gram-positive and Gram-negative vegetative bacteria are susceptible to high-pressure CO2 treat-
ment, with Gram-positive being slightly more resistant to the pressurized CO2 owing to the
thick peptidoglycan layer. Studies conducted by Tortora et al. (2002) and Dillow et al. (1999)
have shown that over 90% of the vegetative bacteria can be completely inactivated by HPCD.
However, for complete sterilization and inactivation of all microorganisms including the most
resistant endospores, at least 10 6cfu/mL (6D reduction) spores must be completely deactivated
in order to claim sterilization.
13.5.1 Factors affecting Microbial Inactivation
Temperature and pressure are the most important factors affecting growth of microorganisms. Each
microorganism has a species-specific maximum temperature over which it is not able to survive. Above
that temperature, proteins denature, cytoplasmic membranes collapse, and the cells lyse, resulting in their
deactivation (Madigan et al., 2002). A wide range of temperatures have been employed for high-pressure
CO2 treatment, from 0°C (Nakamura et al., 1994) to 100°C (Roskey and Sikes, 1994; Sikes and Martin,
1995). The inactivation rate of microorganisms can be increased by repeated release and recharge of pres-
surized CO2 during the treatment to improve transfer of intracellular materials out of the bacterial cells
(Lin et al., 1992, 1993). HPCD is lethal to bacteria, yeast, and fungi at moderate pressure and low tem-
perature, thus preserving more quality attributes of the products than traditional treatments (Ferrentino
et al., 2009; Spilimbergo and Ciola, 2010). Bacteria are more resistant to pressure than to temperature. A
HHP between 100 and 1000 MPa is required to inactivate bacteria (Cheftel, 1995). However, in presence
of high-pressure CO2, the pressure requirement can be lowered below 20 MPa. The highest pressure
in HPCD reported is only 33 MPa to get similar inhibition (Arreola, 1991). As far as the mechanism is
concerned, in HHP, pressure is applied to the food indirectly; however, during HPCD applications, the
high-pressure CO2 is in direct contact with food. Process temperature for HPCD treatment ranges from
20°C up to 60°C while process pressure ranges between 7.0 to 40.06 MPa. The treatment time can range
from about 3 and 9 min to about 120 and 140 min depending on the food that is being processed and the
mode of operation, i.e., continuous, semi-continuous, or batch-type (Thakur et al., 2013).
13.6 ENZYME INaCtIVatION
As highlighted in the previous section, HPCD is an effective non-thermal processing technique for
inactivating deleterious enzymes in liquid as well as solid foods. Indigenous enzymes in food such as
polyphenol oxidase (PPO), pectin methylesterase (PME), and lipoxygenase (LOX) may cause undesir-
able chemical changes in food attributes, manifested by the loss in color, texture, and flavor (Hu et al.,
2013). HPCD treatment by avoiding high temperatures exerts a minimal impact on the nutritional and
sensory properties of foods, with simultaneous extension of shelf life by inhibiting or killing enzymes.
13.6.1 Mechanism of action
All enzymes are proteinaceous in nature, and therefore stable in certain pressure, temperature,
and pH regimes. Invasion by CO2 can cause an imbalance of intra-molecular and solvent-protein
interactions, leading to partial or complete denaturation of the enzyme depending on the severity
of treatment (Ishikawa et al., 1996; Truong et al., 2002; Park et al., 2002; Kincal et al., 2006). The
HPCD treatment may also affect the conformational (tertiary) structure of enzymes (Chen et al.,
1992; Gui et al., 2006a). Even though it is a nonthermal treatment, impact of HPCD on α-helix and
β-sheet structure of protein is more severe than thermal treatment (Ishikawa et al., 1996; Zhou et al.,
2009). This shows that enzymes are more resistant to heat than to pressure. Studies have revealed
conformational changes of tryptophan and tyrosine caused as a result of SC-CO2 application (Gui
et al., 2006a; Zhou et al., 2009). Most of the commercial enzyme preparations express only 90% of
their initial activity at 1 h exposure to 20.3 MPa at 35°C (Taniguchi et al., 1987). Presence of ethanol
and high moisture content has proved to further enhance the severity of the treatment for enzyme
inactivation. Possible mechanisms responsible for enzyme inhibition are discussed below.
13.6.1.1 Inhibitory Effects of Molecular CO2
Higher pressure results in decrease in intermolecular space resulting in simultaneous increase in

density of CO2 molecules. Dense CO2 molecules act like a hydrophobic medium that interacts with
the hydrophobic groups of the protein, compelling them to be exposed as per the theory of “Like
Dissolves Like” (Yoshimura et al., 2001). Exposure to CO2 disintegrates the active site of enzymes
making them non-functional (Clifford and Williams, 2000).
Temporary anaerobic environment generated by CO2 is also responsible for partial or total inacti-
vation of the enzyme. This temporary anaerobic environment gets totally vanished when atmospheric
pressure is retained. This may possibly be the reason behind the reactivation of enzymes when CO2
leaves the matrix in most of the cases. This also explains the potential of CO2 at atmospheric pressure
for inactivation of lipase up to 84% (Fadıloglu and Erkmen, 2002).
Reversibility has been observed in various enzymes such as PME and PPO just after treatment
as well as during storage (Hu et al., 2013). The possible reason of reactivation may be as a result of
restoration of the aerobic condition which was earlier dominated by temporarily generated anaero-
bic environment. However, synergistic effects have also been reported in case of HPCD like HHP
for example, the effect of SC-CO2 extraction conditions (temperature, pressure, extraction time,
and moisture content of the samples) on the myrosinase activity, and glucosinolate hydrolysis in
flaked and whole canola seeds was studied. Combined effects of high temperature (75°C), pressure
(62.1 MPa), and moisture (20% w/w) were necessary to achieve 90% enzyme inactivation in canola
flakes within 3 h (Dunford and Temelli, 1996).
13.6.1.2 The pH Lowering Effect
As stated earlier, the HPCD-induced inactivation of enzymes is based on the hypothesis that
CO2 can dissolve in water, causing a reduction in pH due to formation of carbonic acid that leads
to inactivation of the enzyme. Although carbonic acid is a weak acid (Ka = 4.5 × 10−7 at 25°C),
reversibility and higher concentration of CO2 causes re-formulation of carbonic acid. This frequent
formulation of carbonic acid causes unstable condition for the enzymes. The pH stability range is
different for different enzymes (Balaban et al., 1991; Zhou et al., 2009). Many enzymes involved in
carbohydrate and amino acid metabolism have pH optima in a neutral range (Hutkins and Nannen,
1993) and their activity declines sharply on either side of that optimum. Phase diagram of carbon
dioxide (Figure 13.1) shows that with increasing pressure and temperature, solubility of CO2 gets
enhanced, leading to increased production of acid. This indicates that by increasing pressure, the
deleterious effect of CO2 can be enhanced (without much increase in temperature).
13.6.1.3 Effects of Depressurization
Repeated cycles of increased pressure and depressurization bring about drastic explosion in
the matrix causing the decomposition of the compact enzyme molecules into smaller particles
(Zhou et al., 2009). When dissolved CO2 pressure releases, it loosens the non-covalent-bonded water
molecules associated with enzymes and reduces their activity since total moiety structure of the
enzyme gets disturbed (Fricks et al., 2006).
13.6.1.4 Possible Other Mechanisms
Certain inactivation mechanisms may be specific to enzymes or to food matrix. Oliveira et al. (2006)
observed changes in the shape of lipase as observed under electron micrographs, from a nearly rounded
form with a smooth surface to a somewhat deformed shape. The change in protein (enzyme) structure
leads to deformation in active site and overall destruction in activity. They also reported enhanced
porosity of enzyme particle by HPCD treatment. Aggregation of protein particles due to high pressure
has also been reported as in case of mushroom tyrosinase, which leads to enzyme inhibition (Zhou
et al., 2009). Aggregation along with acid-induced protein coagulation and homogenization effect may
be the principle reasons behind the change in particle size of proteins or enzymes (Zhou et al., 2009).
13.6.2 Factors affecting Enzyme Inhibition
Enzyme inhibition is governed by several factors such as time of exposure, pressure level, tem-
perature, pH of food matrix, the food ingredients, HPCD processing system, processing cycle, etc.
Sensitivity of individual enzymes varies; for example, cysteine arylamidase, α-galactosidase,

α and β-glucosidase, and N-acetyl-β-glucosaminidase were found to be significantly affected by
HPCD, whereas lipase, leucinearylamidase, β-galactosidase, phosphatase (acid and alkaline), and
naphthol-AS-BI-phosphohydrolase were only slightly affected (Hong and Pyun, 2001). A summary
of literature has been detailed below.
13.6.2.1 Effects of Pressure Levels
Theoretically, pressure level and degree of enzyme inhibition are directly related to each other.
For example, inactivation of PPO and PME can be subsequently enhanced as pressure level is
increased (Park et al., 2002). Both enzymes are major problems of fruit processing industries, with
the former associated with browning while the latter associated with decrease in firmness of the
plant cell. Inactivation of both the enzymes in fruits is required to retain firmness, freshness, and
appeal of the fruit for extended period. Gui et al. (2007) concluded that by increasing pressure
from 8 to 30 MPa, PPO activity reduced by 20% in apple juice. In horseradish, red beet, apple
and orange juice, A. niger, and B. subtilis, the respective enzymes such as POD, PPO, PME, acid
protease, and α-amylase follow first-order degradation kinetics in response to differential pressure
(Hu et al., 2013). Ishikawa et al. (1995) observed that alkaline protease and lipase can be completely
inactivated at a pressure above 15 MPa. LOX, which is responsible for beany flavor in soybean, can
be inactivated one fourth of its initial activity by a 15 min exposure at 10.3 MPa at 35°C (Tedjo et al.,
2000). Complete inactivation of the enzyme was observed at 62.1 MPa. This demonstrated a direct
effect of pressure on enzyme activity.
Researchers have shown that HHP treatments below 400 MPa have little effect on the activity of
PME in orange juice (Cano et al., 1997; Basak and Ramaswamy, 1998; Nienaber and Shellhammer,
2001). Since HHP is based on Pascal’s isostatic principle stating that pressure applied to a sam-
ple is transmitted in a uniform and instantaneous manner and Le Chatelier’s principle stating the
phenomena of phase transition, chemical changes which are accompanied by the decrease in volume
are favored by the pressure, so that reactions leading to an increase in volume will tend to be inhibited
by pressure and vice versa (Venugopal et al., 2001). Therefore, in comparison with HPCD treatment,
HHP is ineffective against detrimental enzymes since the applied pressure in HPP is uniformly dis-
tributed. However, in HPCD treatment targeted action leads to change in structure of these enzymes
through one or the other mode of action such as denaturation, aggregation, acid-induced coagulation,
homogenization, particle porosity development, etc. Thus, HPCD and HHP have different influence
on enzyme activity and would appear to operate through different mechanisms. Though pressure
level in HHP is many fold higher then HPCD, but due to different mode of action, more inhibition is
achieved by HPCD. Many enzymes have been reported in literature to be severely affected by HPCD
treatment, but HHP always requires temperature, pH, CO2 treatments (Boff et al., 2003) in the form
of an additional hurdle.
13.6.2.2 Effect of Temperature
As per the concept of temperature coefficient (Q10), severity of any treatment which involves
enzymatic reaction and physiological processes is significantly influenced by temperature; there-
fore, rate of enzyme inhibition gets enhanced by increasing temperature. This may be due to fast
diffusion and more collisions between the applied CO2 and target enzymes (Ishikawa et al., 1995;
Tedjo et al., 2000; Gui et al., 2006a, 2007; Niu et al., 2009). Being a non-thermal treatment, tem-
perature during HPCD treatment needs to be maintained below safe range (~60°C).
Tedjo et al. (2000) have reported 100% reduction in activity of peroxidases and LOX at pres-
sure of 10.3 MPa within a temperature range of 35°C to 55°C. Similar effect of temperature rise has
been observed for PPO and POD inactivation in apple and horseradish (Gui et al., 2006b, 2007).
Pectin methylesterase activity, which is responsible for cloudiness in apple juice, was reduced by
36% with an increase in temperature from 25°C to 55°C (Niu et al., 2009).
13.6.2.3 Effect of Treatment Time
In general, extension of HPCD process time has been shown to decrease the activity of enzymes
(Balaban et al., 1991; Chen et al., 1992; Yoshimura et al., 2002; Gui et al., 2006a; Zhi et al., 2008).
This may be due to induced irreversibility of the treatment. When treatment time increases, enzymes
do not get chance to recover and ultimately get inactivated.
13.6.2.4 Effect of pH, Food Ingredients, and Processing Media
Each enzyme has its specific pH maxima where it exhibits maximal activity as well as pH
minima having opposite impact. If the supercritical stage is supported by pH minima of the food
matrix or similar range of pH, HPCD would have maximum effectiveness for enzyme inhibition.
Sugars, salts, polyvalent ions, and alcohols have shielding effect on protein and protect them
from denaturation, thus, reducing severity of the treatment (Lakshmi and Nandi, 1976; Van den
Broeck et al., 1999). This may be due to colligative effect of some of the previously mentioned com-
ponents which reduce the thermal severity of HPCD. Colligative effect may become more severe
with increase in viscosity as is the case of sucrose above 50% concentration (Tedjo et al., 2000).
In the presence of salt and sugar, water activity of food matrix also gets reduced, which further
decreases the solubility of CO2 and protects enzymes from deterioration.
13.6.2.5 Effect of Microbubbled CO2
Microbubbles (MBs) are very tiny bubbles in liquids, with diameters of the order of micrometres
(Parmar et al., 2013). In HPCD, microbubbling is associated with the large surface area given to
CO2. Microbubbling provides more advantages over conventional HPCD as surface area increases
the chances of collision of CO2 with enzymes or adsorption of CO2 by the enzymes, which results
in overall decrease in activity of enzymes (Ishikawa et al., 1995; Yoshimura et al., 2001). At lower
concentration, temperature, and pressure, more inactivation can be achieved by microbubbled CO2,
which may be due to increase in the surface area.
13.6.2.6 Pressure Cycle
Number of pressure cycles may enhance severity of inhibition of enzymes in certain cases—for
example, the proteinase from Carica papaya and lipase enzyme. However, if the initial optimized
process conditions are adequate for enzyme inhibition, there would be no effect of increasing pres-
sure cycles as in case of stearase and lipase from Aspergillus niger (Oliveira et al., 2006; Steinberger
et al., 1999).
13.7 appLICatION OF hpCD IN BEVEraGE INDUStrY
HPCD has been successfully applied to various beverages, with the studies reporting different
affects due to variation in temperature and pressure of treatment. In particular, the technology is
beneficial for beverages rich in anthocyanins and ascorbic acid that get destroyed during thermal
treatments and thus results in loss of color and functional properties of the food. Table 13.1 gives a
brief on studies conducted on beverages for achieving desired effects.
table 13.1 Studies on the Effect of high-pressure CO2 on Beverage Quality

S. No Beverage processing Conditions Effect references
1. Citrus juice 3000–6000 psi at Debittering; 25% reduction Kimball (1987)
30°C–60°C in limonin content.
2. Cider 250 bar at 40°C with 6, Dealcoholisation Medina and
8 and 10 kg/h Co2 Martinez (1997)
3. Mandarin juice 41.1 MPa for 9 min at 35°C Cloud enhanced by 38%, Lim et al. (2006)
pectin esterase activity
reduced by 7%–51%,
3.47 log reduction in total
aerobic count.
4. Muscadine grape 48.3 MPa at 0% or 15% Co2 Partial PPo inactivation Pozo-insfran et al.
juice (40%); greater (2007)
polyphenolics and
antioxidant capacity
retention.
5. red beet juice 37.5 MPa for peroxidase reduction in PoD and Liu et al. (2008)
(PoD) and 22.5 MPa for PPo activities by ~86%
PPo at 55°C for 60 min and 95%, respectively.
HPCD reduced the
decimal reduction time of
PoD and PPo from
555.56 to 74.63 min and
161.29 to 38.31 min,
respectively.
6. Carrot juice 10–30 MPa for 15–60 min browning degree and pH of Zhou et al. (2009)
at 25°C HPCD-treated carrot
juices decreased, the
cloud and the titratable
acidity increased
significantly.
7. Apple juice 20 MPa with Co2 Aerobic bacteria, yeasts, Liao et al. (2010)
concentration of 4.5%–5.3% and moulds were totally
at 52°C inactivated, the microbial
counts were <10 CFu/mL.
8. Litchi juice 10 MPa at 32°C–52°C for reduction of 4.19 log Li et al. (2012)
5–30 min with mild heat cycles by HPCD + MH at
(MH) and nisin 52°C for 15 min; complete
inactivation of aerobic
bacteria by combined
treatment of HPCD, MH
and nisin.
9. Hibiscus 34.5 MPa, 8% Co2, 6.5 min 5 and 0.85 log reductions ramirez-rodrigues
sabdariffa and 40°C for yeasts and moulds and (2013)
beverage aerobic plate count; no
effect on °brix and pH.
10. Watermelon 30 MPa for 15 min at 50°C the activity of PPo, PoD, Liu et al. (2013)
juice and PMe decreased; pH
value and lycopene
content of treated juice
slightly decreased.
11. Coconut water 120 bar, 40°C, 30 min 5 log reduction of Cappelletti et al.
mesophiles, LAb, yeasts (2015)
and moulds and a 7 log
reduction of the total
coliforms; reductions in
volatile fractions.
A method for simultaneously carbonating and cooling for the production of beverages such as
soft drinks has been patented by Karr (1978). The method involves the injection of liquid CO2 into
the bottom of a carbonating column of liquid. The liquid flows upwardly through the column,
resulting in simultaneous cooling and carbonation.
13.8 EXtraCtION OF NUtraCEUtICaLS
Nutraceuticals are the food components which provide proven health benefits apart from
basic nutrition. They are interchangeably used with functional foods but functional foods gen-
erally do not include dietary supplements. Also, nutraceuticals are more refined and isolated
forms of bioactive compounds. One may understand that foods having nutraceuticals can be
categorized as functional foods. In present days, there is a huge scope of functional foods and
nutraceuticals and people are willing to pay high cost for them. In this direction, isolation,
extraction, characterization, and functional food product development are the research areas
which are now catching huge attention of the researchers. Presently, carotenoids, flavonoids,
and other phenolic compounds, fatty acids and lipids, and other phytochemicals are the major
nutraceuticals gaining attention.
In this context, application of supercritical fluid extraction (SCFE) for the isolation and frac-
tionation of the high-value components is on the rise due to their organic nature and requirement
of high feed-to-solvent extraction ratio. SCFE is a separation process where solid or liquid matter
is treated with a supercritical fluid in order to obtain diffusible compounds from miscella. A super-
critical fluid is above its critical temperature and possesses gas-like viscosity, liquid-like density
and its diffusion ability. SCFE offers a variety of applications due to easily adjustable solubilizing
properties by changing pressure and temperature. In general, temperature and pressure have shown
the biggest influence on the solubility of compounds in supercritical fluids. As per the theory of
“Like Dissolves Like”, pure CO2 can be a solvent of nonpolar molecule, but it cannot dissolve the
polar or water-soluble molecule; therefore, some polar modifier such as methanol, ethanol, etc.
needs to be added to dissolve water-soluble pigments in it. A polar modifier or co-solvent is gen-
erally added with SC-CO2 to enhance solubility. Selection of modifiers requires much attention
as not all modifier may give similar efforts like in case of xanthophyll extraction from Japanese
persimmon, addition of ethanol decreases the overall yield (Takahashi et al., 2006). Sometimes,
even some vegetable oils can act as better modifiers than alcohols like in the case of xanthophylls
extraction from marigold (Gao et al., 2009). Residual effect of polar modifier is also of major con-
cern. Additional setup/reaction/unit operation may be required to keep them separate from major
compounds of interest. Sometimes, as a cost-effective approach under subcritical conditions, a
water system has also proved to be a successful polar modifier like in the case of phenolic com-
pound extraction from rosemary (Crego et al., 2004). Selection of co-solvent also depends on the
solubility of the desired component. The extraction of bioactives is dependent on a number of
variables as discussed in the next section.
13.8.1 Effect of temperature
Temperature has two competing effects on solubility. Increasing the temperature at constant
pressure decreases the density of the solvent. Thus, solubility of the solute is decreased. On the other
hand, by increasing the temperature at constant density, the vapor pressure of the solute is increased
and the solute becomes more soluble in a supercritical fluid. Which of these effects will prevail is
dependent on the properties of the solute.
13.8.2 Effect of pressure
With increasing pressure of a supercritical medium, higher densities are achieved; the higher the
density of the medium, the higher the solubility of the solute (Vazquez et al., 2006). By differentiating
the pressure, various bioactive compounds can be fractionated from the same biological material.
By varying the solubility, different components can be fractionated like at elevated pressures. For
example, water is poorly soluble with SC-CO2, while ethanol is completely soluble. Therefore, two
phases can be made when pressurized CO2 is introduced into a water-ethanol mixture (Nunes et al.,
2007). A scientific database of solvents has been generated by Nunes et al. (2007) on the basis of
solubility which can be referred for selection of solvent systems.
Last but not the least, the extraction rates depend on the morphology of the material and the
location of the solute in plant material since mass transfer depends on particle shape, size, and the
porosity of the solid material. In general, one should reduce the diffusion path and diffusion resis-
tance to get higher yield. Each process parameter is required to be examined and controlled very
precisely. Theoretically, reduction in particle size enhances the extraction rate but simultaneously
chances of agglomeration increase, which reduces the extraction yield (Liu et al., 2011).
13.8.3 Extraction of Carotenoids
Carotenes and xanthophylls are the two major categories of carotenoids. Carotenoids are
well-documented nutraceuticals as well as natural pigments (Britton, 1995). They possess antioxi-
dative potential. To date, carotenoid extraction from various food commodities has been exercised
using SCFE using CO2 as a solvent or chief solvent. As per the results compiled by Oman et al.
(2013), members of carotenoids family that have been extracted from different biological materi-
als such as microalgae, tomatoes, pumpkin, lotus, marigold, palm oil, etc. include astaxanthin,
canthaxanthin, lutein, lycopene, zeaxanthin, β-carotene, β-cryptoxanthin, etc. Freeze drying is rec-
ommended before extraction of carotenoids from the biological material keeping thermo-stability
of nutraceuticals in view. Particle size, solventization power, co-solvent (having different polarity),
flow rates, and flow direction affect the overall yield of carotenoids or other bioactives (Subra et al.,
1998). Extraction of β-carotene and lycopene has been studied from tomato waste by Baysal et al.
(2000) at pressure level of 30 MPa.
13.8.4 Extraction of Flavonoids
Since flavanoids are polar compounds, their separation without polar modifiers is not possible
(Scalia et al., 1999; Antolovich et al., 2000; Bergeron et al., 2005; Chang et al., 2007; Araujo et al.,
2007). To date, anthocyanins, apigenin, artepillin C, baicalin, camellianin A, catechin, cinnamic
acid, gallic acid, ellagic acid, genistein, glabridin, quercetin, resveratol, stilbenes, γ-oryzanol, etc. are
few of the major flavanoids that have been extracted from different biological materials such as
olives, tea, soybean, pigeon pea, rice, grape, black currant, marigold, hops, etc. using SC-CO2 with
ethanol, methanol, water, hexane as co-solvent/polar modifier.
Semenova et al. (1992) conducted one of the first studies related to the use of SC-CO2 coupled
with membrane technology. The first operational supercritical fluid extraction unit was patented
by Robinson and Sims (1996), that involved coupling of SCF and hollow fibre membranes which
is now commercially referred to as “PoroCrit” or as membrane-based dense gas extraction. In the
“PoroCrit” process, an aqueous liquid solution is circulated on one side and on the other side
the extraction solvent, i.e., near-critical or supercritical CO2 is flowing co-directionally. When
the membrane used is hydrophobic, the aqueous solution does not penetrate into the membrane pores.
A meniscus is formed at the mouth of the pores stabilizing a dense gas-liquid interface. The mem-
brane does not play a fundamental separation role; instead, the solute extraction is determined by
the chemical potentials (i.e., equilibrium between the phases) and the membrane allows a controlled
contact among the phases. Hollow fibre membranes (which are used in the “PoroCrit” process)
present contact areas considerably greater than conventional contact systems, making them ideal
for this type of process. The “PoroCrit” process has proved effective in orange aroma extraction
(Sims et al., 1998), vanillin and methyl tert-butyl ether extraction (Sims, 1998; Sims et al., 1998),
caffeine extraction (Shirazian, and Ashrafizadeh, 2010) and as a continuous pasteurization assem-
bly for liquid foods (Sims and Estigarribia, 2002).
13.8.5 Extraction of Flavoring Compounds
Since flavor compounds are thermolabile, their retention can be enhanced when non-thermally
isolated. Being a non-thermal technique, SCFE has huge potential to extract flavor compounds from
substrate. It is a potential alternative to the conventional steam distillation and microwave-assisted
extractions.
Essential oils contain monoterpene and sesquiterpene hydrocarbons and oxygenated com-
pounds (alcohols, aldehydes, ketones, acids, phenols, oxides, lactones, ethers, esters), which are
responsible for the characteristic odors (Capuzzo et al., 2013). They are volatile and often non-polar
in nature. As already discussed, for extraction of polar compounds, it is suggested to use polar
modifier or cosolvent system. Selection of polar modifier should be based on the critical pressure
and temperature. It should be in the range of major solvent so that during desolventization, polar
modifier can also be removed without any extra unit operation and cost. Nitrous oxide has a critical
temperature and pressure of 36.7°C and 7.1 MPa, which is very close to that of CO2 i.e. 31.2°C
and 7.3 MPa, respectively. But due to handling problems, ethanol is a more preferred solvent than
nitrous oxide. Ethane, propane, and dimethyl ether are also often used as co-solvents.
Isolated material must be fractionated before inclusion in food products to avoid impurities
of racemic mixture (Flores et al., 2005) since only the volatile portion is responsible for aroma.
Sometimes terpene creates problem in delivery of essential oil, therefore, deterpination is required,
in that case ethane is a better solvent than supercritical carbon dioxide (Raeissi et al., 2008). As per
the data compiled by Oman et al. (2013), a number of essential oils such as 1,8-cineole, 4-nonanone,
4-terpineol, carvacrol, carvone, caryophyllene, cedrene, chrysanthenol acetate, cinnamonaldehyde,
citral, citronellol, eugenol, eugenyl acetate, geranial, geraniol, germacrone, limonene, linalool, linalyl
acetate, menthol, β-caryophyllene, β-selinene, γ-terpinene, τ-cadinol, etc. have been extracted from
various plant-based biological material using SC-CO2. Many of them have also been extracted with
hydro-distillation as well as with microwave assistance. Various researchers have compared the yield
of essential oils from plant materials using different methods (Wang et al., 2010; Patel et al., 2012).
Yield in terms of percent content varied as per the plant material and type of essential oil. For instance,
yield of myrcene from coriander was the highest when extracted by SC-CO2, however, microwave
assisted extraction was found best in case of fennel. In contrast, content of α-limonene extracted from
Foeniculum vulgare was the lowest using SCFE method in comparison with microwave assisted and
hydro-distillation method. Patel (2013) compared all three methods for the yield of various fractions of
essential oil in seed of Cuminum cyminum, Foeniculum vulgare, and Coriandrum sativum, as well as
the cost effectiveness. Keeping extraction times, yield, processing charges, raw material cost as input
variables; cost of SCFE was the lowest in comparison to hydro-distillation and microwave-assisted
extraction in all three plant materials (taking all fractionable essential oil into account). Generally, for
extraction of α-pinene, D-limonene, cuminal, caryophyllene, β-terpeneol, carinol, geraniol, glyceryl
monooleate, heptdecenal, myrcene, etc., SCFE is a preferred method over microwave assisted and
hydro-distillation extraction. However, SCFE was found ineffective for extraction of myrtinyl acetate,
octadecenoic acid, and hexadecenal.
13.8.6 Encapsulation Using SC-CO2
Encapsulation can be achieved by supercritical fluids based on their properties and their effi-
ciency can be easily changed with variations in pressure and temperature. Carbon dioxide is the
most widely used supercritical fluid because it offers low-temperature processing capabilities
making it highly suitable for heat-sensitive materials. SC-CO2 can be used for encapsulation in three
different ways (Munin and Levy, 2011) as a solvent, an anti-solvent, or a solute. The supercritical
solvent/anti-solvent processing is used when the solute is sparingly soluble in the supercritical fluid.
SC-CO2 behaves as the anti-solvent. SC-CO2 is injected into a pressurized container having extrac-
tion solution and solute to be encapsulated. On introduction into the reaction chamber, SC-CO2 comes
in contact with the solution that dissolves in its phase, thereby, decreasing density and increasing the
solvation power of the organic solvent. At the same time, the solvent evaporates in the supercritical
phase, causing oversaturation of the solution and, thus, precipitating the solute. The precipitated
solute is microencapsulated and the excess solvent can be removed via purge gate. Sosa et al. (2011)
encapsulated green tea polyphenols with a biodegradable polymer (polylactide-polycaprolactone
copolymer or PLC) by co-precipitation using SC-CO2. The solute process utilizes SC-CO2 as a
solute because under pressure it can dissolve in liquids. It is also called as supercritical-assisted
atomization (SAA) process. For encapsulation, a large quantity of SC-CO2 is solubilized in a melted
substance, dissolved, or dispersed in a solvent forming a gas-saturated solution. This solution is
expanded into the atomization chamber through a nozzle to form solid particles or liquid droplets.
The combined action of the cooling mixture and the volume expansion of gas lead to precipitation of
the substance forming microencapsulated particles which are collected in the reactor after depres-
surization (Meterc et al., 2008).
13.9 SUpErCrItICaL FLUID EXtrUSION
Supercritical fluid extrusion (SCFX) is a novel extrusion technology combining the expanding
and plasticizing properties of supercritical carbon dioxide (SC-CO2) as an expanding gas
(Rizvi et al., 1995). Rizvi and Mulvaney (1992) patented the production of highly expanded starch
foam by SCFX. It is a unified process of thermoplastic extrusion employing SC-CO2 as a blowing
agent (Figure 13.2). The SCFX process permits the use of heat-sensitive materials in the formulation
as carbon dioxide is at its supercritical state at approximately ambient temperature (31.1°C) and
thus the need of high temperature during processing is avoided. This novel process can be used for
manufacturing new generation of biomaterials for food as well as for non-food applications.
Figure 13.2 super critical fluid extruder.

The general expansion mechanism of SCFX process as highlighted by Alavi et al. (1999) and
Mulvaney and Rizvi (1993) consists of the following major steps:
1. Development of gas holding matrix by heat-shear treatment

2. SC-CO2 injection into the matrix to create a saturated solution
3. Nuclei formation due to thermodynamic instability by sudden pressure drop at the die
4. Expansion and cell growth at the die exit when pressure drops to atmospheric level
The SCFX process offers the following advantages:
1. It permits the use of heat sensitive materials in the formulation.

2. In-line addition of flavorings, colorants, and bioactive materials can be done.
3. It allows manipulation of in-barrel pressure and temperature during processing to yield different
microcellular, composite (sandwich) structures.
4. It selectively and preferentially deposits bioactives on the inside walls of the porous extrudates.
5. It aids in reduction of melt viscosity and pH (due to large solubility effects).
6. It provides more degrees of freedom to modify the rheological and chemical properties of the
extrudates.
SCFX offers a plethora of applications in food and packaging industry and has proved to be a
versatile technology for multiple end uses. The SCFX process has the potential for improvement
in processing, manufacture, and quality of a range of puffed food products, such as ready-to-eat
cereals, pasta, and confectionery products with improved texture, color and taste. It also offers
improvements to packaging materials due to its plasticizing and microcellular expansion properties.
Some of the food applications are detailed below.
13.9.1 Bakery
SCFX can be used in bakery applications as it leads to aeration and leavening of dough without
addition of any biological (yeast) or chemical (sodium bicarbonate) leavening agent. SCFX allows
for continuous production of yeast-free dough leavened via incorporation of SC-CO2. The incor-
poration of SC-CO2 can ensure continuous aeration and proofing that yields bake-ready dough in
less time. The extruded dough can directly be baked in ovens for manufacture of leavened baked
goods without actual fermentation by yeast or enzymes. Moreover, a uniform cell-size distribution
achieved by SC-CO2 within the dough can improve the bread quality characteristics. This is more
desirable for leavening dough than the conventional means for chemical leavening. The process can
also support multiple ingredients as nutritional needs of yeast are avoided. Extrusion also manipu-
lates structure of starch and thus multigrain systems can also be used to prepare healthy baked
goods.
Khanitta and Rizvi (2009) successfully developed premixed dough leavened by 1% (feed basis)
SC-CO2 injection in a twin-screw extruder at 37°C at specific mechanical energy input (18 kWh/ton).
The dough was baked using a combination of vacuum and conventional baking to reduce the baking
time. This process allowed a continuous system of dough development for production of consistent
ready-to-bake dough. The breads were developed in a shorter duration and had uniform quality attri-
butes equivalent to commercial products. Simultaneously, the process eliminated ethanol emissions.
Hicsasmaz et al. (2003) concluded that the SCFX-leavened dough has rheological properties
comparable to the commercial dough prepared by mixing, yeast fermentation, and proofing. SCFX
produces the leavened dough in about 2 min, eliminating the need for proofing and holding time and
can be done in a continuous-line processing facility. The continuous nature of this entire process is
an attractive asset and provides a definite economic advantage. SCFX also offers similar advantages
in other baked products such as buns, muffins, doughnuts, croissants, etc.
13.9.2 Extruded Snacks
Snacks and convenience foods form the most progressive food segment of present times. Fried
starch-based snacks are frequently portrayed as less than desirable foods because of their high
caloric content and low nutrient density. Increasing concerns about health has led to increasing
demand for new healthy snacks as an alternative for fried starch-based snacks with low nutrient
density. Extruded snack foods offer advantages over fried snacks in terms of health and also the pro-
cess allows fortification and nutrient enrichment in products. Extrusion cooking is widely practiced
in cereal-based snack products because of its versatility in terms of ingredient acceptability, shape,
size, and characteristics of end products. In conventional steam-based extrusion, water behaves
both as a plasticizer for melt formation of the feed ingredients and as a blowing agent for expansion
to yield a puffed product (Wang et al., 2005). When the melt passes through the extruder die, it is
exposed to a sudden pressure drop resulting in water vapor nuclei generation. The vapor in nuclei
evaporates instantaneously due to release of pressure and leaves an expanded solid porous structure.
Steam-based extrusion (single or twin screw) generally requires low moisture (18%–28%), high
temperature (120°C–180°C), and high shear conditions to achieve good expansion. These harsh
operating conditions often lead to nutrient damage and excessive dextrin formation causing hard-
ness in the product and also limits effective utilization of heat- and shear-sensitive ingredients such
as proteins, vitamins, and certain flavors.
As supercritical carbon dioxide can deliver certain flavors, an expanded snack not only with high
nutrient density and unique texture, but also with encapsulated flavors can be produced using this tech-
nology and can be marketed as a novel healthy snack. SCFX chips were developed by Cho and Rizvi
(2008) having uniform cellular microstructure (uniform porosity) that cannot be obtained using con-
ventional steam-based extrusion. As SCFX extrusion is conducted at low temperature (~60°C–80°C)
and low shear conditions compared with conventional steam-based extrusion, nutritional loss is mini-
mized. Moreover, SC-CO2-expanded extrudates show predominantly homogeneous closed cell struc-
tures (microcellular nuclei) and non-porous surface that facilitates flavor encapsulation and provides
better textural control over products manufactured using steam-based extrusion (Alavi et al., 1999).
Post-extrusion processes such as baking and frying not only impart desirable flavor and appearance
to the SCFX product but also generate further expansion as SCFX extrudates demonstrate a time-
delayed expansion until the structure is completely set (Alavi et al., 1999; Chen et al., 2002). The
SCFX process allows control of the average cell size of extrudates to range from about 50–250 μ and
the volumetric expansion of final products by manipulating process parameters such as die dimension,
SC-CO2 concentration, and SC-CO2 residence time (Alavi and Rizvi, 2005; Winoto, 2005).
The technology to prepare ready-to-cook convenience foods such as noodles, pasta, macaroni,
vermicelli, etc. is known as cold extrusion where the feed is formed into dough in extruder and
shaped to final product. These products can be instantized by precooking them to gelatinize the
starch followed by re-drying before packaging. SC-CO2 processing can alter the microstructure of
traditionally prepared pasta product to obtain quick cooking pasta. The process can be used to inject
SC-CO2 into an uncooked semolina pasta using a short-barrel version of the SCFX process. This
process is aided by the fact that pasta extrusion is an inherently high-pressure and low-temperature
process so as not to denature the flour proteins. The SCFX process can be used to infuse flavor and
microstructure chocolate and confectionery items. Microstructuring adds to mouthfeel of the con-
fectionary products by decreasing their density.
13.9.3 texturization Using SCFX
Texturization is a process by which food materials can be given a specific texture. It is majorly
done for vegetable proteins so as to imitate meat proteins and for dairy and other ingredients to
improve their functional properties such as binding, foaming, gelling, and emulsification.
The extrusion process has been applied to vegetable proteins to imitate animal proteins,
replacing the traditional technique with a continuous process and developing food products with
new textural characteristics. SCFX offers sub-100°C expansion of vegetable proteins using direct
SC-CO2 injection. The delicate balance of temperature and shear during SCFX processing permits
controlled modification of protein conformation. Addition of SC-CO2 provides additional acidic
environment and serves as a surface modifier of the protein matrix that markedly improves binding,
gelling, and emulsifying characteristics. SCFX in presence of certain chemicals or modifying
agents (“reactive SCFX”) such as mineral salts (CaCl2 and NaCl) create highly alkaline or acidic
environment which in combination with controlled shear and heat favorably alter the gelling and
functional properties of proteins (Manoi and Rizvi, 2009). Structural changes in SCFX proteins
due to denaturation and polymerization lead to an increased surface hydrophobicity and molecular
flexibility. Emulsifying activity is enhanced by denaturation as a result of increased hydrophobic
interactions between adjacent protein molecules at the interface and structural rigidity (Kato and
Nakai, 1980; Matsudomi et al., 1985; Mitidieri and Wagner, 2002; Guilmineau and Kulozik, 2007).
Manoi and Rizvi (2009) found that “reactive SCFX” contributed approximately 258 and
275,000 times higher apparent viscosity and elastic modulus than the unprocessed WPC. Texturized
WPC samples produced under extremely acidic (pH 2.89) and alkaline (pH 8.16) conditions with
SC-CO2 exhibited high stability of rheological properties over a wide temperature range (25°C–85°C).
13.9.4 polymer Foaming
Foamed polymers are an attractive packaging material as they provide cushioning and resis-
tance to packed product amongst other benefits. Foamed plastics are cost effective compared to
conventional plastics owing to their low mass per unit volume. They also provide thermal and sound
insulation to packed product. Foamed polymers due to their porosity fare poorly in terms of gas and
moisture barrier properties. Conventional foams are prepared by extrusion technology that entraps
air at the die exit in small cells (nucleation) followed by large volume expansion of nuclei and simul-
taneously prevents coalescence of cells by freezing the extrudate surface. More recently, the previ-
ously described surface cooling method was used in an extrusion-based open-cell foaming process
with supercritical CO2 for thermoplastic polymers such as polystyrene (PS) and polycarbonate (PC).
The effects of CO2 content, surface quenching (chilling), die geometry, and temperature lead to dif-
ference cellular structure, densities, and strengths of foamed polymeric material. SCFX has been
used successfully to produce biopolymeric foams with bubble size in the range of 50–200 μ and
bubble density to the order of 106 per cm3 (Alavi et al., 2002).
The application of SC-CO2 in extrusion technology offers multiple advantages and creates a
range of novel products and processes. In food and packaging applications the ability of supercritical
CO2 to behave as a blowing and plasticizing agent leads to improved puffed and expanded products,
uniform flavor and ingredient distribution, texturization of proteins, and manufacture of microcel-
lular polymeric foams. Thus, incorporation of SC-CO2 adds to the versatility of extrusion technology
and forms an important application of utilization of super critical CO2 in the food industry.
Supercritical CO2 is the most widely used solvent in SCFE technology owing to its easy availability
and low cost. The extraction is facilitated at near room temperatures and relatively low pressures.
13.10 EFFECt ON FOOD QUaLItY attrIBUtES
HPCD is believed to have minimal effect on food quality attributes. Researchers have reported
least influence of the process on the bioactive compounds such as ascorbic acid and carotenoids pres-
ent in fruit juices (Boff et al., 2003; Zhou et al., 2009) owing to the exclusion of oxygen from the food
matrix and the lowering of pH. Although instrumental color of the product in terms of L* and a* values
are slightly affected by the HPCD treatment (Park et al., 2002; Kincal et al., 2006), no difference in
sensory property of HPCD-treated and control samples during low-temperature storage for 14 days
has been observed. Kincal et al. (2006) stated that the threshold value for the panellists for the feel of
“carbonation effect” in the product was at 230 ppm CO2. Precipitation of calcium and calcium carbon-
ate and reduced particle size as a result of high pressure has been shown to result in enhanced cloudi-
ness in citrus, carrot, and peach juices, which further delays the destabilization of colloidal suspensions
(Park et al., 2002; Boff et al., 2003; Lim et al., 2006; Kincal et al., 2006; Zhou et al., 2009). The HPCD
resulted in a significant reduction in the degree of browning in juices during storage at 4°C as compared
to untreated sample, indicating that the process could effectively prevent the browning of apple juice
(Gui et al., 2006b).
In the case of fresh-cut fruits and vegetables, HPCD was found to cause adverse effect on fruit firm-
ness and texture (Valverde et al., 2010; Spilimbergo et al., 2013). A study conducted by Messens et al.
in 1997 concluded that HPCD could induce protein denaturation and aggregation in the meat. HPCD
treatment at 7.4 MPa at 31.1°C for 10 min did not affect the pH, tenderness, or water-holding capacity
of the treated meat. Chao et al. (1991) reported that treatment of meat with SC-CO2 solubilized and
removed up to 36.9% of the cholesterol and 71.2% of total lipids in the meat during the process.
13.11 appLICatION OF hpCD FOr WatEr DISINFECtION
Water quality in terms of microbiological quality needs to be maintained if it has to be used as

a food ingredient. Also, the waste water generated needs to be treated so as to reduce its microbial
load before it is discharged as an effluent. Various approaches for water disinfection involve the
use of chlorine, ozone, UV radiations, and certain chemicals. Lately, the application of pressurized
CO2 for disinfection of water has come in the limelight. The first study for water disinfection was
carried out by Kobayashi et al. (2007). Before that, the studies on application of HPCD dealt with
dried foods and beverages. Evidence suggests that when experiments using CO2 under pressure
were conducted with high water content or high moisture, the presence of water made cell walls
more permeable to CO2, with higher diffusivity, higher solubility, and lower viscosity than in dried
media (Kamihira et al., 1987; Haas et al., 1989; Lin et al., 1993).
Ishikawa et al. (1997) introduced the inactivation of bacterial spores in an aqueous medium by
the MB method. They compared the inactivation efficiency of the supercritical CO2 treatment with
that by thermal treatment at 100°C. B. polymyxa spore population was reduced by 0.85 log cycles
after heating at 100°C for 60 min whereas the supercritical carbon dioxide treatment of B. polymyxa
spores at 31°C and 30 MPa for 15 min led to a 6-log reduction. In another study conducted by
Vo et al. (2013), DNA-based viruses showed higher susceptibility to CO2 MBs as compared to
RNA-based viruses at pressures of about 0.7 MPa. They also observed a synergistic effect of pres-
surized CO2 with the increasing temperatures to bring about a 2-log reduction of E. coli at 13 ±
0.2°C for 25 min and also absence of E. coli at 26.6 ± 4°C for 20 min.
13.12 DISINFECtION OF StOrED GraINS
Application of pressurized CO2 has also been tested to combat pests, insects, and mites in
stored food grains. The hazardous effect of fumigants and synthetic chemicals applied for insect
disinfestation of stored grains and the subsequent ban on them has led researchers towards finding
alternative solutions for killing of the storage pests. Apart from the control of pests, the other main
issues in the search for new fumigants for foodstuffs are the effect on quality, registration issues,
and mode and ease of application (Desmarchelier, 1998). Recently, interest has grown in the area
of use of pressurized CO2 for the disinfestations of stored grains and dried products owing to its
nontoxic nature and zero residue effect. The potential for use of HPCD started as early as 1973
by Mitsura et al. (1973), who reported the potent effect against the grain mite Tyrophagus putres-
centiae. They recommended treatment by CO2 at 1600 kPa bar pressure for 10 min at 40% CO2.
Subsequently, Stahl et al. (1985) used the technology for removal of storage pests in medicinal
plants. The first batch-type commercial plant came up in Germany in the year 1988 for disinfesta-
tion of spices, herbs, and pet foods (Pohlen et al., 1989). Utilizing this technology, Nakakita et al.
(2001) have further developed a “disinfester” with rotary valves that can treat large amount of grains
with pressurized CO2 in a continuous mode. Sen et al. (2009) have also reported a lethal impact of
high-pressure CO2 against pests in dried figs.
Lethality of the HPCD varies with the life-cycle stage of the pest with the eggs being the most
resistant, requiring higher pressures and longer exposure times for 100% removal, followed by
larvae, adults, and pupae. Lethality of the technology has been reported to be indirectly propor-
tional to the treatment temperature (Ulrichs et al., 1997). Prozell and Reichmuth (2001) proposed
that it is necessary to identify the pest and classify its susceptibility to HPCD.
The efficiency of CO2 depends on the concentration of gas, duration of exposure, temperature,
product’s moisture content, and insect species and life stage (Jay, 1984). Applying a modified
atmosphere under high pressure (25 bars) proved to be effective in shortening the exposure period
required for complete insect mortality, as reported by Rajendran (2001). CO2 treatment under high
pressure provides rapid control of insects, but its use is limited to high-value crops because large
quantities of gas are needed and the chambers are expensive (Adler et al., 2000).
HPCD treatment thus requires shorter duration for lethal exposure and can be highly promising
for quarantine treatment and rapid disinfestations of valuable products.
Pressure of up to 40–50 bar and ambient temperature (Gerard et al., 1988; Quirin, 1988) was
found to be successful in killing pests such as Lasioderma serricorne (Cigarette beetle), Tribolium
confusum, Stegobium paniceum (drugstore beetle), Plodiainter punctella, and Acarussiro
(flour mite). Mortality of different pest-insects was found to depend on treatment time and pressure
of CO2 along with the stage of insect’s development. A cent per cent mortality is guaranteed after
a treatment time in the range of 5–120 min, depending on the pressure applied and the stage of the
insect’s development (Quirin, 1988).
A patent has also been granted in 1993 for use of liquid CO2 as a fumigant fluid (US Patent
5394643 A) to Hans-Bernd Schmittmann (1993). Carbon dioxide, in a liquid form under high
pressure, is introduced into the storage area through underground tunnels. Liquid CO2 penetrates
into the cavities and produces low temperatures, at which time the pests are killed by exposure to
sub-zero temperatures. Further, CO2 gas is formed in situ by sublimation in the immediate vicinity
of the pests present in cracks and crevices, consequently suffocating them. No toxic traces are left
behind and the process is harmless to the environment and ecology.
13.13 LIMItatIONS OF thE tEChNOLOGY
Although the HPCD process offers many advantages, it has certain limitations also.
1. Long treatment time is a potential problem of the CO2 sterilization technique. Even though a high
degree of deactivation of spores has been realized, this usually requires more than 10 h, which is not
competitive with the average time of 10–15 min for commercial sterilization.
2. Direct contact of CO2 with the food has to be ensured; therefore, efficiency of the process in solid
food matrices needs extensive research.
3. Foods processed by HPCD are not expected to be commercially sterile and, therefore, require
refrigerated storage immediately after processing.
4. The process is not fit for soft foods or leafy vegetables.

5. It may lead to generation of pressure-induced mutants.
6. The chief hazard in its use is asphyxiation. Any leakage in the plant may lead to death of the
personnel.
13.14 rEGULatOrY aSpECtS
An important issue in the application of HPCD technology in the food preservation/processing

industry is the regulatory approval, which emphasizes microbiological and toxicological safety of
food products. The major problem that novel, nonthermal preservation methods such as HPCD
face in gaining widespread acceptance is that thermal processes are established procedures that
render safe food with high quality and nutritional value in large volumes at very low processing
costs. Generally, high-pressure preservation processes reduce the microbial load to the same level
achieved by traditional technologies, while delivering higher quality products. However, in order to
implement this new technology in the food industry, there is an increased need to understand the
mechanism and kinetics of pressure-induced degradation/denaturation and inactivation of several
food compounds (e.g., nutrients, proteins, microorganisms, enzymes) and the way in which the
degradation/denaturation/inactivation is affected by other parameters such as temperature and pH.
Theoretically, repeated exposure of any stress may generate mutants that for adaptability may have
potential risk to the safety of foods. However, practically only Hong et al. (1999) have observed
some changes in sensitivity towards pressure in L. plantarum cells when repeated cycles of pressure
were given. Therefore, extensive research work is required in this direction to validate the safety
of foods.
Further, inactivation of microorganisms and enzymes are not precisely understood, like in the
case of thermal processing. There may be chances of interaction of food ingredients with HPCD
that leads to some untraceable but toxic compounds. Lack of studies on diffusion mechanisms,
interaction of HPCD with other food components, mechanistic studies, and mathematical model-
ling of the inactivation kinetics are the major points of study.
The HPCD process has versatile usage but, to date, only a few applications have been
commercialized. In addition to food preservation, extraction, fractionating, and product development,
HPCD offers novel usages such as stabilization and sustained release of bioactives and drugs, water
disinfection and stored grains disinfestations, and packaging. However, safety evaluation and interac-
tion of HPCD with other food components need to be focused on simultaneously to assess the risk
associated with treated foods. There is an urgent need for unbiased scientific studies to get convincing
data to find the risk and hazards generated within the food matrix after the treatment.
rEFErENCES
Adler, C., Corinth, H.G., and Reichmuth, C. (2000). Modified atmospheres. In: Subramanyam, B.H., Hagstrum,
D.W. (Eds.), Alternatives to Pesticides in Stored product IPM. Kluwer Academic Publishers, Norwell,
MA, pp. 105–146.
Alavi, S.H., Gogoi, B.K., Khan, M., Bowman, B.J., and Rizvi, S.S.H. (1999). Structural properties of
protein-stabilized starch-based supercritical fluid extrudates. Food Res. Int. 32: 107–118.
Alavi, S.H., and Rizvi, S.S.H. (2005). Strategies for enhancing expansion in starch-based microcellular foams
produced by supercritical fluid extrusion. Int. J. Food Prop. 8: 23–234.
Alavi, S.H., Rizvi, S.S.H., and Harriott, P. (2002). Process dynamics of starch-based microcellular foams
produced by supercritical fluid extrusion. I: Model development. Food Res. Int. 36: 309–319.
Antolovich, M., Prenzler, P., Robards, K., and Ryan, D. (2000). Sample preparation in the determination of
phenolic compounds in fruits, Analyst 125: 989–1009.
Araujo, J.M.A., Silva, M.V., and Chaves, J.B.P. (2007). Supercritical fluid extraction of daidzein and genistein
isoflavones from soybean hypocotyl after hydrolysis with endogenous beta-glucosidases, Food Chem.
105: 266–272.
Arreola, A.G., Balaban, M.O., Wei, C.I., Peplow, A., Marshall, M., and Cornell, J. (1991). Effect of supercritical
carbon dioxide on microbial populations in single strength orange juice. J. Food Qual. 14: 275.
Balaban, M.O., Arreola, A.G., and Marshall, M. (1991). Inactivation of pectinesterase in orange juice by
supercritical carbon dioxide. J. Food Sci. 56, 743–746.
Balaban, M.O., and Duong, T. (2014). Dense phase carbon dioxide research: Current focus and directions.
Agric. Agric. Sci. Procedia 2: 2–9.
Basak, S., and Ramaswamy, H.S. (1998). Ultra high-pressure treatment of orange juice: A kinetic study on
inactivation of pectin methylesterase. Food Res. Int. 29: 601–607.
Baysal, T., Ersus, S., and Starmans, D.A.J. (2000). Supercritical CO2 extraction of beta-carotene and lycopene
from tomato paste waste. J. Agric. Food. Chem. 48: 5507–5511.
Bergeron, C., Gafner, S., Clausen, E., and Carrier, D.J. (2005). Comparison of the chemical composition of
extracts from Scutellaria lateriflora using accelerated solvent extraction and supercritical fluid extrac-
tion versus standard hot water or 70% ethanol extraction. J. Agric. Food. Chem. 53: 3076–3080.
Boff, J.M., Truong, T.T., Min, D.B., and Shellhammer, T.H. (2003). Effect of thermal processing and carbon
dioxide-assisted high-pressure processing on pectinmethylesterase and chemical changes in orange juice.
J. Food Sci. 68: 1179–1184.
Britton, G. (1995). Structure and properties of carotenoids in relation to function. J. FASEB, 9: 1551–1558.
Calvo, L., and Torres, E. (2010). Microbial inactivation of paprika using high-pressure CO2. J. Supercrit.
Fluids 52: 134–141.
Cano, M.P., Hernandez, A., and De Ancos, B. (1997). High pressure and temperature effects on enzyme inac-
tivation in strawberry and orange products. J. Food Sci. 62: 85–88.
Cappelletti, M., Ferrentino, G., Endrizzi, I., Aprea, E., Betta, E., Corollaro, M.L., Charles, M., Gasperi, F.,
and Spilimbergo, S. (2015). High pressure carbon dioxide pasteurization of coconut water: A sport drink
with high nutritional and sensory quality. J. Food Eng. 145: 73–81.
Capuzzo, A., Maffei, M.E., and Occhipinti, A. (2013). Supercritical fluid extraction of plant flavors and fra-
grances. Molecules 18: 7194–7238.
Chang, Y., Liu, B., and Shen, B. (2007). Orthogonal array design for the optimization of supercritical fluid
extraction of balicalin from roots of Scutellaria baicalensis.Georgi. J. Sep. Sci. 30: 1568–1574.
Chao, R.R., Mulvaney, S.J., Bailey, M.E., and Fernando, L.N. (1991). Supercritical CO2 conditions affecting
extraction of lipid and cholesterol from ground beef. J. Food Sci. 56: 183–187.
Cheftel, J.C. (1995). Review: High-pressure, microbial inactivation and food preservation. Int. J. Food Sci.
Technol. 1: 75–90.
Chen, J.S., Balaban, M.O., Wei, C., Marshall, M.R., and Hsu, W.Y. (1992). Inactivation of polyphenol oxidase
by high-pressure carbon dioxide. J. Agric. Food Chem. 40: 2345–2349.
Chen, K.H., Dogan, E., and Rizvi, S.S.H. (2002). Supercritical fluid extrusion of masabased snack chips.
Cereal Foods World. 47: 44–51.
Cho, K.Y., and Rizvi, S.S.H. (2008). The time-delayed expansion profile of supercritical fluid extrudates. Food
Res Int. 41: 31–42.
Clifford, A.A., and Williams, J.R. (2000). Introduction to supercritical fluids and their applications. In: Supercritical
Fluid: Methods and Protocols. pp. 1–16. Clifford A.A., Williams J.R., (Eds.), Humana Press, Totowa, NJ.
Cooper, A.I. (2001). Recent developments in materials synthesis and processing using supercritical CO2. Adv.
Mater. 13: 1111.
Crego, A., Ibanez, E., Garcia, E., de Pablos, R.R., Senorans, F.J., Reglero, G., and Cifuentes, A. (2004).
Capillary electrophoresis separation of rosemary antioxidants from subcritical water extracts. Eur.
Food Res. Technol. 219: 549–555.
Damar, S., and Balaban, M.O. (2006). Review of dense phase CO2 technology: Microbial and enzyme inacti-
vation, and effects on food quality. J. Food Sci. 71: R1–R11.
Daniels, J.A., Krishnamurthi, R., and Rizvi, S.S.H. (1985). A review of effects of carbon dioxide on microbial
growth and food quality. J. Food Protec. 48: 532–537.
Desmarchelier, J.M. (1998). Potential new fumigants. In: Stored Grain in Australia: Proceedings of the
Australian Postharvest Technical Conference, 26–29 May Canberra, Australia, pp. 133–137.
Dillow, A.K., Dehghani, F., Hrkach, J.S., Foster, N.R., and Langer, R. (1999). Bacterial inactivation by using
near- and supercritical carbon dioxide. Proceedings of the National Academy of Sciences of the United
States of America 96: 10344–10348.
Dunford, N.T., and Temelli, F. (1996). Effect of supercritical CO2 on myrosinase activity and glucosinolate
degradation in canola. J. Agric. Food Chem. 44: 2372–2376.
Fadıloglu, S., and Erkmen, O. (2002). Inactivation of lipase by carbon dioxide under atmospheric pressure.
J. Food Eng. 52: 331–335.
Ferrentino, G., Plaza, M.L., Ramirez-Rodrigues, M., Ferrari, G., and Balaban, M.O. (2009). Effects of dense
phase carbon dioxide pasteurization on the physical and quality attributes of a red grapefruit juice.
J. Food Sci. 74: E333–E341.
Ferrentino, G., and Spilimbergo, S. (2011). High pressure carbon dioxide pasteurization of solid foods: Current
knowledge and future outlooks. Trends Food Sci. Technol. 22: 427–441.
Flores, G., Blanch, G.P., Del Castillo, M.L. R., and Herraiz, M. (2005). Enantiomeric composition studies in
Lavandula species using supercritical fluids. J. Sep. Sci. 28: 2333–2338.
Foster, J.W., Cowan, R.M., and Maag, T.A. (1962). Rupture of bacteria by explosive decompression. J. Bacteriol.
83: 330–334.
Fraser, D. (1951). Bursting bacteria by release of gas pressure. Nature 167: 33–34.
Fricks, A.T., Souza, D.P.B., Oestreicher, E.G., Antunes, O.A.C., Girardi, J.S., Oliveira, D., and Dariva, C.
(2006). Evaluation of radish (Raphanus sativus L.) peroxidase activity after high-pressure treatment
with carbon dioxide. J. Supercrit. Fluids 38: 347–353.
Gao, Y.X., Nagy, B., Li, X., Simandi, B., and Wang, Q. (2009). Supercritical CO2 extraction of lutein esters
from marigold (Tagetes erecta L.) enhanced by ultrasound. J. Supercrit. Fluids 49: 345–350.
Garcia-Gonzalez, L., Geeraerd, A.H., Spilimbergo, S., Elst, K., Van Ginneken, L., and Debevere, J. (2007).
High pressure carbon dioxide inactivation of microorganisms in foods: The past, the present and future.
Inter. J. Food Microbiol. 117: 1–28.
Gerard, D., Kraus, J., and Quirin, K.W. (1988). Rückstandsfreie Druckentwesung mit natürlicher Kohlensäure
[Residue free pressure disinfestation with natural carbon dioxide]. Gordian 88: 90–94.
Gui, F., Chen, F., Wu, J., Wang, Z., Liao, X., and Hu, X. (2006a). Inactivation and structural change of horse-
radish peroxidase treated with supercritical carbon dioxide. Food Chem. 97: 480–489.
Gui, F., Wu, J., Chen, F., Liao, X., Hu, X., Zhang, Z., and Wang, Z. (2006b). Change of polyphenol oxidase
activity, color, and browning degree during storage of cloudy apple juice treated by supercritical carbon
dioxide. Eur. Food Res. Technol. 223: 427–432.
Gui, F., Wu, J., Chen, F., Liao, X., Hu, X., Zhang, Z., and Wang, Z. (2007). Inactivation of polyphenol oxidases
in cloudy apple juice exposed to supercritical carbon dioxide. Food Chem. 100: 1678–1685.
Guilmineau F., and Kulozik U. (2007) Influence of a thermal treatment on the functionality of hen’s egg yolk
in mayonnaise. J. Food Engg., 78: 648–654.
Haas, G.J., Prescott, H.E., Dudley, E., Dik, R., Hintlian, C., and Keane, L. (1989). Inactivation of microorgan-
isms by carbon dioxide under pressure. J. Food Safety 9: 253–265.
Haas, G.J., Prescott, H.E., Dudley, E., Dik, R., Hintlian, C., and Keane, L. (1989). Inactivation of microorgan-
isms by carbon dioxide under pressure. J. Food Safety 9: 253–265.
Hicsasmaz, Z., Dogan, E., Chu, C., and Rizvi, S.S.H. (2003). Leavened dough processing by supercritical fluid
extrusion (SCFX). J. Agric. Food Chem. 51: 6191–6197.
Hong, S.I., Park, W.S., and Pyun, Y.R. (1997). Inactivation of Lactobacillus sp. from kimchi by high pressure
carbon dioxide. Lebensmittel-Wissenschaftund-Technologies, 30: 681–685.
Hong, S.I., and Pyun, Y.R. (1999). Inactivation kinetics of Lactobacillus plantarum by high pressure carbon
dioxide. J. Food Sci. 64: 728–733.
Hong, S.I., and Pyunb, Y.R. (2001). Membrane damage and enzyme inactivation of Lactobacillus plantarum
by high pressure CO2 treatment. Int. J. Food Microbiol. 63: 19–28.
Hu, W., Zhou, L., Xu, Z., Zhang, Y., and Liao, X. (2013). Enzyme inactivation in food processing using high
pressure carbon dioxide technology. Crit. Rev. Food Sci. Nutr. 53: 145–161.
Hutkins, R.W., and Nannen, N.L. (1993). pH homeostasis in lactic acid bacteria. J. Dairy Sci. 76:
2354–2365.
Ishikawa, H., Shimoda, M., Kawano, T., and Osajima, Y. (1995). Inactivation of enzymes in an aqueous
solution by micro-bubbles of supercritical carbon dioxide. Biosci. Biotech. Biochem. 59: 628–631.
Ishikawa, H., Shimoda, M., Tamaya, K., Yonekura, A., Kawano, T., and Osajima, Y. (1997). Inactivation of
Bacillus spores by the supercritical carbon dioxide micro-bubble method. Biosci. Biotechnol. Biochem.
61: 1022–1023.
Ishikawa, H., Shimoda, M., Yonekura, A., and Osajima, Y. (1996). Inactivation of enzymes and decomposi-
tion of helix structure by supercritical carbon dioxide microbubble method. J. Agric. Food Chem. 44:
2646–2649.
Jay, E.G. (1984). Imperfections in our current knowledge of insect biology as related to their responses to
controlled atmospheres. In: Controlled Atmosphere and Fumigation in Grain Storages. B.E. Ripp,
H.J. Banks, E.J. Bond, D.J. Calverley, E.G. Jay and S. Navarro (Eds.), Elsevier, Amsterdam, the
Netherlands, pp. 493–508.
Kamihira, M., Taniguchi, M., and Kobayashi, T. (1987). Sterilization of microorganisms with supercritical
carbon dioxide. Agric. Biol. Chem. 51: 407–412.
Karr, FA. (1978). Liquid carbon dioxide carbonation method. US 4068010 A.
Kato, A., and Nakai, S. (1980). Hydrophobicity determined by fluorescence probe method and its correlation
with surface properties of proteins. Biochemicaet Biophysica Acta 624: 13–20.
Kauffman, F.L., Shank, J.L., and Urbain, W.M. (1969). Irradiation with CO2 under pressure, US 3483005.
Khanitta, M., and Rizvi, S.S.H. (2009). Physicochemical changes in whey protein concentrate texturized by
reactive supercritical fluid extrusion. J. Food Eng. 95: 627–635.
Kimball, D.A. (1987). Debittering of citrus juices using supercritical carbon dioxide. J. Food Sci. 52: 481–482.
Kincal D., Hill W.S., and Balaban M. (2006). A continuous high-pressure carbon dioxide system for cloud and
quality retention in orange juice. J. Food Sci. 71: C338–C344.
Kincal, D., Hill, W.S., Balaban, M.O., and Marshall, M.R. (2006). A continuous high pressure carbon dioxide
system for microbial reduction in orange juice. J. Food Sci. 70: M249–M254.
Kobayashi, F., Hayata, Y., Kohara, K., Muto, N., and Osajima, Y. (2007). Application of supercritical CO2
bubbling to inactivate E. coli and coliform bacteria in drinking water. Food Sci. Technol. Res. 13: 20–22.
Lakshmi, T.S., and Nandi, P.K. (1976). Effects of sugar solutions on the activity coefficients of aromatic amino
acids and their N-acetyl ethyl esters. J. Phys. Chem. 80: 249–252.
Liao, H.M., Zhang, L.Y., Hu, X.S., and Liao, X.J. (2010). Effect of high pressure CO2 and mild heat processing
on natural microorganisms in apple juice. Int. J. Food Microbiol. 137: 81–87.
Lim, S.L., Yagiz, Y., and Balaben, M.O. (2006). Continuous high pressure CO2 processing of mandarin juice.
Food Sci. Biotechnol. 15: 13–18.
Lin, H.M., Yang, Z.Y., and Chen, L.F. (1992). Inactivation of Saccharomyces cerevisiae by supercritical and
subcritical carbon dioxide. Biotechnol. Prog. 8: 458–461.
Lin, H.M., Yang, Z.Y., and Chen, L.F. (1993). Inactivation of Leuconostoc dextranicum with carbon dioxide
under pressure. Chem. Eng. J. 52: B29–B34.
Liu, J., Lin, S.Y., Wang, Z.Z., Wang, C.N., Wang, E.L., Zhang, Y., and Liu, J.B. (2011). Supercritical fluid
extraction of flavonoids from Maydis stigma and its nitrite-scavenging ability. Food Bioprod. Process
89: 333–339.
Liu, X., Gao, Y., Peng, X., Yang, B., Xu, H., and Zhao, J. (2008). Inactivation of peroxidase and polyphe-
nol oxidase in red beet (Beta vulgaris L.) extract with high pressure carbon dioxide. Innov. Food Sci.
Emerging Technol. 9: 24–31.
Liu, Y., Hu, X.S., Zhao, X.Y., and Zhang, C. (2013). Inactivation of polyphenol oxidase from watermelon juice
by high pressure carbon dioxide treatment. J. Food Sci. Technol. 50: 317–324.
Li, H., Zhao, L., Wu, L., Zhang, Y., and Liao, X. (2012). Inactivation of natural microorganisms in litchi juice
by high-pressure carbon dioxide combined with mild heat and nisin. Food Microbiol. 30: 139–145.
Madigan, M.T., Martinko, J.M., and Parker, J. (2002). Brock Biology of Microorganisms. Prentice Hall, Upper
Saddle River, NJ.
Manoi, K., and Rizvi, S.S.H. (2009). Rheological characterizations of texturized whey protein concentrate-
based powders produced by reactive supercritical fluid extrusion. Food Res. Int. 41: 786–796.
Matsudomi, N., Sasaki, T., Kato, A., and Kobayashi, K. (1985). Conformational changes and functional prop-
erties of acid-modified soy protein. Agric. Biol. Chem. 49: 1251–1256.
McHugh, M., and Krukonis, V. (1993). Introduction Supercritical Fluid Extraction Butterwoth-Heinemann.
Newton, MA, 1–16.
Medina, I., and Martínez, J.L. (1997). Dealcoholisation of cider by supercritical extraction with carbon
dioxide. J Chem. Technol. Biotechnol. 8: 14–18.
Mertens, B., and Knorr, D. (1992). Developments of nonthermal processes for food preservation. Food Tech.
46: 124–133.
Meterc, D., Petermann, M., and Weidner, E. (2008). Drying of aqueous green tea extracts using a supercritical
fluid spray process. J. Supercrit. Fluids 45: 253–259.
Mitidieri, F.E., and Wagner, J.R. (2002). Coalescence of o/w emulsions stabilized by whey and isolate soybean
proteins. Influence of thermal denaturation, salt addition and competitive interfacial adsorption. Food
Res. Int. 35: 547–557.
Mitsura, A., Amano, R., and Tanabe, H. (1973). The acaricidal effects of compressed gas treatments on the
grain mite, Tyrophagus putrescentiae. Shokuhin Eiseigaku Zashi 14: 511–515.
Moshashaee, S., Bisrat, M., Forbes, R.T., Nyqvist, H., and York, P. (2000). Supercritical fluid processing of
proteins. Part I. Lysozyme precipitation from organic solution. Eur. J. Pharm. Sci. 11: 239.
Mulvaney, S.J., and Rizvi, S.S.H. (1993). Extrusion processing with supercritical fluids. Food Technol. 47:
74–82.
Munin, A., and Lévy, F.E. (2011). Encapsulation of natural polyphenolic compounds: A Review. Pharmaceutics
3: 793–829.
Nakakita, H., Ikenaga, H., and Takahashi, K. (2001). Disinfestation of stored grains using high-pressure
carbon dioxide In: Proceedings of the International Conference on Controlled Atmosphere and
Fumigation in Stored Products. Donahaye, E.J., Navarro, S., and Leesch J.G. (Eds.), Fresno, CA. 29th
October–3rd November. 2000, Executive Printing Services, Clovis, CA, pp. 421–430.
Nakamura, K., Enomoto, A., Fukushima, H., Nagai, K., and Hakoda, M. (1994). Disruption of microbial-cells
by the flash discharge of high-pressure carbon dioxide. Biosci. Biotechnol.Biochem. 58: 1297.
Nienaber, U., and Shellhammer, T.H. (2001). High-pressure processing of orange juice: Kinetics of pectin-
methylesterase inactivation. J. Food Sci. 66: 328–331.
Niu, S., Xu, Z., Fang, Y., Zhang, L., Yang, Y., Liao, X., and Hu, X. (2009). Comparative study on cloudy apple
juice qualities from apple slices treated by high pressure carbon dioxide and mild heat. Innov. Food Sci.
Emerg. 11: 91–97.
Nunes, A.V.M., Matias, A.A., Da Ponte, M.N., and Duarte, C.M.M. (2007). Quaternary phase equilibria for
SC-CO2 plus biophenolic compound plus water plus ethanol. J. Chem. Eng. Data 52: 244–247.
Oliveira, D., Feihrmann, A.C., Rubira, A.F., Kunita, M.H., Dariva, C., and Oliveira, J.V. (2006). Assessment of
two immobilized lipases activity treated in compressed fluids. J. Supercrit. Fluids 38: 373–382.
Oman, M., Skerget, M., and Knez, Z. (2013). Application of supercritical fluid extraction for the separation of nutra-
ceuticals and other phytochemicals from plant material. Macedonian J. Chem. Chem. Eng. 32: 183–226.
Park, S.J., Lee, J.I., and Park, J. (2002). Effects of a combined process of high-pressure carbon dioxide and
high hydrostatic pressure on the quality of carrot juice. J. Food Sci. 67: 1827–1834.
Parmar, R., and Majumder, S.K. (2013). Microbubble generation and microbubble-aided transport process
intensification-A state-of-the-art report. Chemical Engineering and Processing: Process Intensification
64: 79–97.
Patel, J.B. (2013). Comparison of the conventional and novel extraction methods Chapter 9: http://shodhganga.
inflibnet.ac.in/bitstream/10603/27156/18/18_chapter%209.pdf. Accessed on March 23, 2017.
Patel, J.B., Patel, B., Patel, R.K., and Patel, B.H. (2012). Comparative evaluation of extraction methods for
extraction of essential oil from Foeniculum vulgare. J. Pharma. Sci. Bioscientific Res. 2: 176–178.
Pohlen, W., Rau G., and Finkenzeller, E. (1989). Erste praktische Erfahrungen mit einem Verfahren zur
Druckentwesung mit Kohlendioxid [First practical experiences with carbon dioxide under pressure for
pest control]. Pharm. Ind. 8: 917–918.
Pozo-Insfran, D., Murat O. Balaban, M.O., and Talcott, S.T. (2007). Inactivation of polyphenol oxidase in
muscadine grape juice by dense phase-CO2 processing. Food Res. Int. 40: 894–899.
Prozell, S., and Reichmuth, C. (2001). Carbon dioxide under high pressure for stored-product protection in
temperate climates. In: Proceedings of the International Conference on Controlled Atmosphere and
Fumigation in Stored Products. Donahaye, E.J., Navarro, S., and Leesch J.G. (Eds.), Fresno, CA.
29th October–3rd November. 2000 Executive Printing Services, Clovis, CA, pp. 719–725.
Quirin, K.W., Stahl, E., and Gerard, D. (1988). Dense Gases Fur Extraction and Refining. Berlin, Germany:
Springer Verlag. pp. 218–223.
Raeissi, S., Diaz, S., Espinosa, S., Peters, C.J., and Brignole, E.A. (2008). Ethane as an alternative solvent for
supercritical extraction of orange peel oils. J. Supercrit. Fluids 45: 306–313.
Rajendran, S. (2001). Alternatives to methyl bromide as fumigants for stored food commodities. Pesticide
Outlook 12: 249–253.
Ramírez-Rodrigues, M.M., Plaza, M.L., Ferrentino, G., Balaban, M.O., Corcuera, J.I.R., and Marshall, M.R.
(2013). Effect of dense phase carbon dioxide processing on microbial stability and physicochemical
attributes of hibiscus sabdariffa beverage. J. Food Process Eng. 6: 125–133.
Rizvi, S.S.H., and Mulvaney, S.J. (1992). Extrusion processing with supercritical fluids. US Patent 5120559.
Rizvi, S.S.H., Mulvaney, S.J., and Sokhey, A.S. (1995). The combined application of supercritical fluid and
extrusion technology. Trends Food Sci. Technol. 6: 232–240.
Robinson, J.R., and Sims, M. (1996). Method and system for extracting a solute from a fluid using dense gas
and a porous membrane, U.S. Patent 5,490, 8840.
Roskey, C.T., and Sikes, A. (1994). Effect of hyperbaric carbon dioxide on spores and vegetative cells of
Bacillus stearothermophilus, NATICK/TR-94/019.
Scalia, S., Giuffreda, L., and Pallado, P. (1999). Analytical and preparative supercritical fluid extraction
of chamomile flowers and its comparison with conventional methods. J. Pharm. Biomed. Anal. 21:
549–558.
Schmittmann, H.-B. (1993). Fumigant fluid Patent US 5394643 A.
Semenova, S.I., Ohya, H., Higashijima, T., and Negishi, Y. (1992). Separation of supercritical CO2 and ethanol
mixtures with an asymmetric polyimide membrane. J. Membr. Sci. 74: 131–139.
Sen, F., Meyvaci, K.B., Aksoy, U., Emekci, M., and Ferizli, A.G. (2009). Effects of the post-harvest application
of methyl bromide alternatives on storage pests and quality of dried fig. Turk. J. Agric. For. 33: 403–412.
Shirazian, S., and Ashrafizadeh, S.N. (2010). Mass transfer simulation of caffeine extraction by subcritical
CO2 in a hollow-fiber membrane contactor. Solv. Extr. Ion Exch. 28: 267–286.
Sikes, A., and Martin, C. (1995). Control of thermophilic spore activity with pressurized CO2 and egg-white
lysozyme, Report, NATICK/TR-95/020.
Sims, M. (1998). Porocritical fluid extraction from liquids using near-critical fluids. Membr. Technol. 97:
11–12.
Sims, M., and Estigarribia, E. (2002). Continuous sterilization of aqueous pumpable food using high pres-
sure carbon dioxide, in Fourth International Symposium on High Pressure Process Technology and
Chemical Engineering, September 22–25, Venice, Italy.
Sims, M., McGovern, E., and Robinson, J.R. (1998). Porocritical fluid extraction application: Continuous
pilot extraction of natural products from liquids with near-critical fluids, Paper presented at the Fifth
Meeting on Supercritical Fluids, Materials and Natural Processing, Nice, France.
Sosa, M.V., Rodríguez-Rojo, S., Mattea, F., Cismondi, M., and Cocero, M.J. (2011). Green tea encapsulation by
means of high pressure antisolvent coprecipitation. J. Supercrit. Fluids 56: 304–311.
Spilimbergo, S., and Ciola, L. (2010). Supercritical CO2 and N2O pasteurisation of peach and kiwi juice. Int. J.
Food Sci. Technol. 45: 1619–1625.
Spilimbergo, S., Komes, D., Vojvodic, A., Levaj, B., and Ferrentino, G. (2013). High pressure carbon dioxide
pasteurization of fresh-cut carrot. J. Supercrit. Fluids 79: 92–100.
Stahl, E., Rau, G., and Adolphi, H. (1985). Entwesung von drogen durch kohlendioxid-druckbehandlung
(PEX-Verfahren) [Disinfestation of drugs with carbon dioxide treatment under pressure (PEX-Method)].
Pharm. Ind. 47, 528–530.
Steinberger, D., Gamse, T., and Marr, R. (1999). Enzyme inactivation and prepurification effects of supercriti-
cal carbon dioxide. In: Proceedings of Fifth Conference on Supercritical Fluids and their Applications,
Italy, Europe. Vol. 339.
Subra, P., Castellani, S., Jestin, P., and Aoufi, A. (1998). Extraction of beta-carotene with supercritical
fluids – experiments and modeling. J. Supercrit. Fluids 12: 261–269.
Takahashi, M., Watanabe, H., Kikkawa, J., Ota, M., Watanabe, M., Sato, Y., Inomata, H., and Sato, N. (2006).
Carotenoids extraction from Japanese persimmon (Hachiyakaki) peels by supercritical CO2 with etha-
nol. Anal. Sci. 22: 1441–1447.
Taniguchi, M., Kamihara, M., and Kobayashi, T. (1987). Effect of treatment with supercritical carbon dioxide
on enzymatic activity. Agric. Biol. Chem. 51: 593–594.
Tedjo, W., Eshtiaghi, M.N., and Knorr, D. (2000). Impact of supercritical carbon dioxide and high pressure on
lipoxygenase and peroxidase activity. J. Food Sci. 65: 1284–1287.
Thakur, M., Blessing, A.J, and Singh, K. (2013). High pressure carbon dioxide–a non-thermal food processing
technique for inactivation of micro-organisms. Asian J. Bio Sci. 8: 267–275.
Tortora, G.J., Funke, B.R., and Case, C.L. (2002). Microbiology: An Introduction, Pearson Education, Boston, MA.
Truong, T.T., Boff, J.M., Min, D.B., and Shellhammer, T.H. (2002). Effects of carbon dioxide in high-pressure
processing on pectinmethylesterase in single-strength orange juice. J. Food Sci. 67: 3058–3062.
Ulrichs, C., Reichmuth, Ch., and Rassmann, W. (1997). Carbon dioxide under high pressure to control the
tobacco beetle Lasioderma serricorne. In: Proceedings of the International Conference on Controlled
Atmosphere and Fumigation in Stored Products, Donahaye, E.J., Navarro, S., and Varnava, A. (Eds.),
April 21–26, 1996, Printco Ltd., Nicosia, Cyprus, 335–341.
Valley, G., and Rettger, L.F. (1927). The influence of carbon dioxide on bacteria. J. Bacter. 14: 101–137.
Valverde, M.T., Marín-Iniesta, F., and Calvo, L. (2010). Inactivation of Saccharomyces cerevisiae in confer-
ence pear with high pressure carbon dioxide and effects on pear quality. J. Food Eng. 98: 421–428.
Van den Broeck, I., Ludikhuyze, L.R., Van Loey, A.M., Weemanes, C.A., and Hendrickx, M.E. (1999).
Thermal and combined pressure-temperature inactivation of orange pectinesterase: Influence of pH and
additives. J. Agric. Food Chem. 47: 2950–2958.
Vazquez, L., Torres, C.F., Fornari, T., Grigelmo, N., Senorans, F.J., and Reglero, G. (2006). Supercritical
fluid extract ion of minor lipids from pretreated sunflower oil deodorizer distillates. Eur. J. Lipid Sci.
Technol. 108: 659–665.
Venugopal, V., Kamal, A.S., and Bongirwar, D.R. (2001). Processing of foods using high hydrostatic pressure.
Ind. Food Industry 20: 65–69.
Vo, H.T., Imai, T., Ho, T.T., Dang, T.L.T., and Hoang, S.A. (2015). Potential application of high pressure
carbon dioxide in treated wastewater and water disinfection: Recent overview and further trends.
J. Environ. Sci. S36: 38–47.
Wang, H.W., Liu, Y.Q., Wei, S.L., Yan, Z.J., and Lu, K. (2010). Comparison of microwave-assisted and con-
ventional hydrodistillation in the extraction of essential oils from mango (Mangifera indica L.) flowers.
Molecules 15: 7715–7723.
Wang, L., Ganjyal, G., Jones, D., Weller, C., and Hanna, M. (2005). Modeling of bubble growth dynamics and
non isothermal expansion in starch-based foams during extrusion. Adv. Polymer Technol. 24: 29–45.
Winoto, C.W. (2005). Process parameters and their influence on supercritical fluid extrudate properties.
Master thesis, Cornell University, Ithaca, NY.
Yoshimura, T., Furutera, M., Shimoda, M., Ishikawa, M., Miyake, M., Matsumoto, K., Osajima, Y., and
Hayakawa, I. (2002). Inactivation efficiency of enzymes in buffered system by continuous method with
microbubbles of supercritical carbon dioxide. J. Food Sci. 67: 3227–3231.
Yoshimura, T., Shimoda, M., and Ishikawa, H. (2001). Inactivation kinetics of enzymes by using continuous
treatment with microbubbles of supercritical carbon dioxide. J. Food Sci. 66: 694–697.
Zhi, X., Zhang, Y., Hu, X., Wu, J., and Liao, X. (2008). Inactivation of apple pectin methylesterase induced by
dense phase carbon dioxide. J. Agric. Food Chem. 56: 5394–400.
Zhou, L., Wang, Y., Hu, X., and Liao, X. (2009). Effect of high pressure carbon dioxide on the quality of carrot
juice. Innov. Food Sci. Emerging Technol. 10: 321–327.
ChaptEr 14
pulsed Magnetic Field processing of Foods
K. Chitravathi and O. P. Chauhan
CONtENtS
14.1 Introduction ........................................................................................................................ 261

14.2 Basic Concepts of Magnetic Fields .................................................................................... 265
14.3 Pulsed Magnetic Field Technology ....................................................................................266
14.3.1 Types of Magnetic Fields .....................................................................................266
14.3.1.1 Static Magnetic Fields and Oscillating/Pulse Magnetic Fields ..........266
14.3.1.2 Ultra-high Magnetic Fields ................................................................. 267
14.3.2 Applications of Magnetic Field in Food Preservation.......................................... 267
14.3.2.1 Effects on Microorganisms and Enzymes .......................................... 267
14.3.2.2 Pasteurization ...................................................................................... 272
14.3.2.3 Sterilization ......................................................................................... 272
14.3.2.4 Magnetic Freezing............................................................................... 275
14.3.2.5 Yeast under High-Gradient Magnetic Field ........................................ 275
14.3.2.6 Isolation and Separation of Protein by Magnetic Technique .............. 279
14.3.2.7 Magnetic Resonance Imaging ............................................................. 279
14.3.3 Application of Magnetic Fields in Quality Control of Food Products ................. 279
14.4 Conclusions and Future Trends ..........................................................................................280
References ......................................................................................................................................280
14.1 INtrODUCtION
The food industry is continuously capitalizing for new preservation techniques owing to the
increase in consumer demand to replace conventional food preservation techniques for foods in
order to reduce carbon footprint of the processes involved in food processing and preservation
for fresh-like quality, natural in taste, nutrient-rich, and safe consumption of foods (Mertens and
Knorr, 1992; Pothakamury et al., 1993). Traditional food processing and preservation methods
261
rely on the use of heat for the inactivation of microorganisms and enzymes responsible for food
spoilage. Although thermal treatment of food extends the shelf life of given products, some
undesirable changes often occur during thermal process. The extent of damage will depend on
the specific food product. The undesirable changes frequently include browning or darkening of
the food, development of “cooked” flavors, degradation of vitamins, loss of texture, etc. However,
nowadays consumers have apparently resigned themselves to eating such products (Sloan, 1999).
Nonthermal technologies are currently undergoing extensive research and the aim is to develop
food processing techniques that will result in products with improved quality. In contrast, to
the traditional thermal processes, these emerging technologies such as high-pressure process-
ing (HPP), pulsed electric field (PEF) technology, pulsed light, ultrasonication, pulsed mag-
netic field (PMF), electron beam, plasma processing, ozone, etc. are predominantly reliant on
enhancing microbial food safety and quality, without compromising the nutritional, functional,
and sensory characteristics of foods and have created an increasing worldwide interest in low
temperature innovative processes for food preservation (Busta et al., 2000; Harte, 2001). The
new processing techniques are mostly employed in the liquid-packed foods when compared to
solid foods. Since the nonthermal methods are used for bulk quantities of foods, these methods
of food preservation are mainly used in the large-scale production. The cost of equipment used
in the nonthermal processing is high when compared to equipment used in thermal process-
ing. After minimizing the investment costs of nonthermal processing methods, it can also be
employed in small-scale industries. Although thermal preservation provides safer food, there
exists loss of food properties like nutrients and sensory attributes. The main objectives of new
techniques are to retain the nutrients and sensory properties and to increase the shelf life without
any adverse effect on its quality. Processing techniques used for the particular products should
be optimized. The selection of particular preservation method for the particular food product is
based on the criteria such as cost of production, scale of production, type of product (i.e., milk,
meat, poultry, fruits, or vegetables), shelf life, and end-product usage (i.e., ready-to-eat or ready-
to-cook product). The nonthermal techniques are recently used for all the food products for shelf
life extension (Mathavi et al., 2013). Table 14.1 gives an overview of the principle and applica-
tions of various nonthermal technologies.
During recent years, the use of PMF technology has attracted much interest as a promising tool
for food processing and preservation and for the creation of new types of food products (Farkas,
2007). The main objective of PMF preservation is to increase the shelf life by reducing the microbial
load and also the water activity. Application of magnetic fields (MFs) to process food products is
one of the novel nonthermal processing methods, and very limited studies have been carried out on
the application and commercialization of this technology. Research on the biological effects of MFs
dates back to as early as 1938 (Kimball, 1938). However, the use of MFs as a nonthermal technol-
ogy for food preservation was first proposed in 1985, when a U.S. patent was granted to Hofmann
(1985). The process could produce fresh-like attributes of foods by retaining thermolabile nutrients,
reduced energy requirements for processing, inactivation of microorganisms, and, in some cases,
the potential treatment of foods inside flexible packages. Application of MFs in the food fermenta-
tion industry could be a major success by controlling cellular growth and inhibition (Ahmed and
Ramaswamy, 2007).
table 14.1 New Non-thermal Food processing Methods
Mechanism of
process principle Description Critical Factors Inactivation Status
HPP HPP subjects foods, with or without Liquid/solid foods, with/without Pressure, time at Denaturation of in use since 1990
packaging, to pressures between packaging (100–800 MPa, pressure, temperature enzymes, proteins; (Japan, usA, France,
100 and 1000 MPa or below 0°C to >100°C, from a (including adiabatic breakdown of spain) Current
80,000–130,000 lb (1–20 min) few seconds to over 20 min heating), pH, biological membranes; pressure processes
instantaneously and composition cellular mass transfer include batch and
uniformly throughout a mass affected semi-continuous
of food independent of size, systems
shape, and food composition
PeF PeF processing involves the High-voltage pulses to foods electric field intensity, Most theories studied Different laboratory-and
application of pulses of high between two electrodes pulse width, treatment are electrical pilot-scale treatment
voltage (20–80 kV per cm) to foods (<1 s; 20–80 kV/cm; time, temperature, breakdown and chambers designed
PuLseD MAGnetiC FieLD ProCessinG oF FooDs
placed between 2 electrodes. exponentially decaying, pulse wave shapes, electroporation and used for foods,
square wave, bipolar, or type, concentration and only two industrial-
oscillatory pulses at ambient, growth stage of scale PeF systems
sub-ambient, or above microorganism, pH, available
ambient temperature) antimicrobials,
conductivity and
medium ionic strength
uV-light/ High-intensity flashes of broad uV radiant exposure, at least transmissivity of the DnA mutations induced used for disinfection of
pulsed spectrum white light pulsed several 400 J/m2. product, the geometry, by DnA absorption of water supplies and
light times from a high-power xenon intense and short-duration the power, wavelength the uV light food contact surfaces
lamp for about 0.1–3 ms per some pulses of broad spectrum and arrangement of
sources or about 100 μs to 10 ms (ultraviolet to the near light source(s), the
per other sources inactivates infrared region) product flow profile
microbes with remarkable rapidity
and effectiveness.
(Continued)
263
264
table 14.1 (Continued) New Non-thermal Food processing Methods
Mechanism of
process principle Description Critical Factors Inactivation Status
ultrasound energy generated by high-frequency energy generated by sound the heterogeneous and intracellular cavitation Combination with e.g.,
sound waves 0.1–20 MHz waves of 20,000 Hz or more protective nature of (micro-mechanical heat, pressure has
food (e.g., inclusion of shocks that disrupt certain potential
particulates) severely cellular structural and
curtails the singular functional components
use of ultrasound for up cell lysis)
preservation
PMF Food is subjected to a high-intensity, subjecting food sealed in Consistent results Controversial results on Application at the
moderate frequency MF in the plastic bags to 1–100 oMF concerning the efficacy effects of MFs on moment not
range of 5–50 t and 5–500 kHz pulses (5–500 kHz, of this method are microbial populations considered
frequencies to destroy or inactivate 0°C–50°C, 25–100 ms) needed
microorganisms in a mainly
non-electrically conductive
environment.
Source: butz, P., and tauscher, P., Food Res. Int., 35, 279–284, 2002.
PuLseD MAGnetiC FieLD ProCessinG oF FooDs 265
14.2 BaSIC CONCEptS OF MaGNEtIC FIELDS
The phenomenon of magnetism was discovered by the Greeks as early as 800 B.C., when it
was first noticed that certain stones attracted pieces of iron. In 1600, William Gilbert suggested
that because compass needles orient in preferred directions, the earth is actually a large permanent
magnet. However, it was not until 1819 that the relationship between magnetism and electricity was
discovered by Danish scientist Hans Oersted. Shortly after, Andre Ampere suggested that electric
currents of molecular size were responsible for all magnetic phenomena, an idea which is now the
basis for the modern theory of magnetism (Serway, 1982). Magnetism is a phenomenon by which
materials exert an attractive or repulsive force on other materials. The origin of magnetism lies in
the orbital and spin motions of electrons and how the electrons interact with each other (Ahmed and
Ramaswamy, 2007).
A MF is also defined as a region of space in which a magnetic body is capable of magnetizing
the surrounding bodies. The MF vector B (also called magnetic induction or magnetic flux density)
is measured in Weber per square meter (Wb/m2), also called Tesla (T). However, in practice, the
cgs unit for MF, called Gauss (G), is often used. One Tesla equals 10,000 Gauss. The magnetic
force is given by F = qv.ΧB, where q is a charged particle moving at a velocity v and Χ indicates
the cross product. B is defined at some point in space in terms of the magnetic force that would be
exerted on a test object, in this case on a charged particle. It must be noted that if q is positive, then
F is in the direction of v Χ B, and if q is negative, the direction of F is opposite to that of v Χ B.
The magnitude of the magnetic force has the value F = qv sin ϴ, where ϴ is the angle between
v and B. Hence, when ϴ is 0° or 180°, v is parallel to B, and F will be zero. On the other hand,
when v is perpendicular to B (ϴ = 90°), F will have its maximum value (Barbosa-Canovas et al.,
2001) (Figure 14.1).
Figure 14.1 (a) Helical movement by a charged particle in a uniform MF b when the velocity vector v makes an
arbitrary angle with b and the (b) circular path followed by the charged particle when v is normal
to b. (From barbosa Canovas, G.V. et al., Magnetic fields as a potential non-thermal technology
for the inactivation of microorganisms, in: Control of Foodborne Microorganisms, pp. 399–418,
Juneja, V. K., and sofos, J. n., eds., Marcel Dekker, new york, 2001.)
14.3 pULSED MaGNEtIC FIELD tEChNOLOGY
The PMF technology is considered as one of the most bolstering technologies to food processing
(Mertens and Knorr, 1992; Wouters and Smelt, 1997). Previous developments of directional MFs
began in 1936, when Davis and Rawls (1974) first discovered that the north and the south magnetic
pole fields each exhibited unique properties with respect to their effects on various forms of life.
This discovery was slow to be accepted because of investigations by others who confused Davis’s
directional polarities MFs with those which are simultaneously emitted by horseshoe-type mag-
nets and by AC-powered electromagnets. Davis and Rawls (1974) presented numerous examples of
enhancement to seeds and various types of cellular growth.
14.3.1 types of Magnetic Fields
Superconducting magnets are a good choice for high-intensity MF generation since they avoid
heating effects. MF generation with coils of superconducting metal has been used by various
researchers (Ahmed and Ramaswamy, 2007). Metals behave as superconductors (electricity passes
through the wire with no resistance) in liquid helium environment (∆268.9°C). This generates a MF
of about 2 T, which is about 40,000 times stronger than the earth’s MF (0.0005 T). The liquid helium
is insulated with liquid nitrogen, which helps to reduce the loss of helium from the magnets. The
magnets need to be filled with liquid nitrogen regularly (weekly), and with liquid helium about once
a month, which makes this type of magnet expensive to run. However, it has also some limitation
in field intensity, with maximum limits of about 20 T. A hybrid magnet contains a superconducting
magnetic coil and water-cooled magnetic coil that can produce 15–30 T. MF above 30 T has been
generated in a pulsed form by supplying current of 40 kA for short time period (Date, 1983).
The application of high-intensity MF is one of the latest innovations in the nonthermal process-
ing of foods. Materials have different responses towards MFs and when the magnetization is equal
along the three orthogonal axes of x, y, and z, the particle is defined to have isotropic susceptibility.
If the magnetization is unequal along the x, y, and z axes, the magnetic susceptibility is known as
anisotropic susceptibility. Carbon atoms as such show isotropic susceptibility, whereas two carbon
atoms bonded together exhibit anisotropic susceptibility. Most of the organic compounds are dia-
magnetic in nature during which the MF applied causes lesser intensity of magnetization under
normal conditions compared with the induction in vacuum. Usually applied MFs could be either
static or oscillating types. In the case of oscillating/pulse magnetic field (OMF) applied in the form
of pulses, it causes reversion in the change with each pulse and intensity of each pulse decreases
with time to about 10 percent of initial intensity (Martin et al., 1997).
Generation of high-intensity MF is usually carried out by supplying current to electrical coils.
The inactivation of microorganisms usually requires magnetic flux densities from 5–50 T. OMFs of
this density can be generated by (a) superconducting coils, (b) coils that produce DC fields, and (c)
coils energized by the discharge of energy stored in the capacitors. Usually OMF circuits use capaci-
tor banks for storage of energy. A capacitor is charged from a voltage source. An oscillating current is
generated between the plates of the capacitor and the same generate OMF. The frequency of the MF
is determined by the capacitance of the capacitor and also the resistance and inductance of the coil.
As the current changes direction, the MF changes polarity. For generating a MF of 70 T, a capacitor
with stored energy of 1.25 MJ is required (Pothakamury et al., 1993). Many foods have significant
electric resistivity greater than 10–25 ohms per cm, which is conducive for the application of OMFs.
14.3.1.1 Static Magnetic Fields and Oscillating/Pulse Magnetic Fields
Hofmann (1985) observed the inactivation of microorganisms exposed to a flux density greater than
2 T, a single pulse with flux density between 5–50 T and frequency of 5–500 kHz reduces the number
of microorganisms by at least 2 log cycles. Metal packages as such cannot be used in MFs. Food pres-
ervation by application of MF is safe to perform. The high-intensity MF exists only within the coil and
immediate vicinity; thus, an operator positioned at a reasonable distance from the coil is out of danger.
The MFs are classified according to their relative strength as low or high intensity, according to the
variation of intensity over space as homogeneous or heterogeneous, and over time as static magnetic
fields (SMFs), OMFs, and ultra-high magnetic fields. Low-intensity MFs show strength in the order
of 1/10th of a Gauss, while high-intensity fields show strength in the order of thousands of Gauss or
greater (Barbosa Canovas et al., 2001). Homogeneous MFs have intensities that are constant over space.
On the other hand, in heterogeneous MFs, the intensity varies across space. The intensity of static
fields remains constant over time, whereas the intensity of pulsed fields varies over time. According
to previous definitions (Figure 14.1) for homogeneous MFs, when v is normal to B, a charged particle
q will move in a circular path, whereas in other orientations between v and B, the charge will move
in a helical path. When charged particles move in heterogeneous fields, the motion is rather complex
(Barbosa-Canovas et al., 2001). SMFs show a constant strength over time and are generated by perma-
nent magnets or direct current electromagnets. An OMF is applied in the form of constant amplitude
or decaying amplitude sinusoidal waves and exhibits an intensity gradient over time depending on the
nature of the magnet. OMFs are generated by alternate current electromagnets within pulsed fields, and
the field intensity alters periodically depending on the frequency and type of wave generating from the
electric current in the magnet (Pothakamury et al., 1993). The MFs may be homogeneous or heteroge-
neous. In a homogeneous MF, the field intensity B is uniform in the area enclosed by the MF coil, while
in the case of heterogeneous the intensity decreases as the distance from the centre of the coil increases.
OMF is applied in form of pulses which reverse the charge for each pulse, and the intensity of each
pulse decreases with time to about 10% of the initial intensity (Barbosa-Canovas et al., 2001). SMFs and
OMFs have been explored for their potential as microbial inactivation methods.
14.3.1.2 Ultra-high Magnetic Fields
The generation of higher field (in mega Gauss ranges) is still difficult because of the coil destruc-
tion by the higher electromagnetic (EM) forces. Miura et al. (1991) have developed ultrahigh MFs in
the mega Gauss range using EM flux compression technique and direct fast current discharge into
a single turn coil. The novel processes can generate 500–1000 T field strength. Scientists from the
National High Magnetic Field Laboratory, University of Florida, have recently developed one super-
conducting magnet, which stands 16 ft tall and weighs more than 15 t. Now, with its commissioning,
scientists from around the world will be able to expand the horizons of scientific investigation using
nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) technologies. The bore
is the space within the magnet that holds the sample being tested. The bore size of this magnet
(105 mm) makes it particularly useful for scientific research. The 900 MHz magnet delivers 21 T
MFs that vary by less than 0.0000002 T over a volume roughly equal to the size of a small orange,
an accomplishment unrivalled anywhere else in the world (Ahmed and Ramaswamy, 2007).
14.3.2 applications of Magnetic Field in Food preservation
14.3.2.1 Effects on Microorganisms and Enzymes
Generally, MFs influence the direction of migration and alter the growth and reproduction of
microorganisms. SMFs and OMFs have been explored for their potentials as microbial inactiva-
tion methods. The inactivation of microorganisms requires magnetic flux densities of 5–10 Teslas
(T) and frequency of 5–500 kHz. MFs up to 3 T can be generated by inserting an iron core in the
coil, above 3 T air-core solenoids are used as iron core shows magnetic saturation. Super con-
ducting magnets can generate high intensities around 20 T without any joule heating, which is a
main difficulty in generating high-intensity MFs. MFs with flux densities of about 15–30 T can
be obtained using hybrid magnet, which is a combination of superconducting magnetic coil with
a water-cooled magnetic coil (Rubin et al., 1983). The coil uses the energy stores in the capacitor
bank, which is charged from a voltage source. When the switch is closed, completing the circuit that
includes the capacitor and the coil, an oscillating current is generated between the plates of capaci-
tor resulting in OMF. As the current changes its direction the MF changes its polarity.
Low intensity MFs like the earth’s MF gives direction to the migration of microorganisms.
Budding was inhibited in cultures when subjecting to MFs due to translocation of paramagnetic free
radicals to a region where field intensity is greatest, resulting in metabolic interruptions. Biological
membranes exhibit strong orientations like parallel or perpendicular to the applied MFs because of
intrinsic anisotropic nature of bio-molecules resulting in resonance of peptide bond. There are two
popular mechanisms for microbial inactivation by MFs.
The ion cyclotron resonance model: At cyclotron resonance where gyrofrequency equals frequency
of applied MFs, energy is transferred to the ions from fields resulting in ionic drifts characterized by
increase in transport of ions like Ca2+ across the cell membrane. Therefore, ions transmit the effects
of MFs from the interaction sites to other tissues and organs like cytoskeleton, nuclear membrane,
chromosomes, and protein molecules (Liboff, 1985).
The ion parametric resonance model: The IPR model predicts ion response correlated with the
changes in experimentally controlled variables, namely magnetic flux densities of the AC and DC
MFs. It also helps in predicting the response of specific ions when the frequency of the MFs corre-
sponds with the ICR conditions. It suggests that during exposure to MFs at ion resonance frequency,
the interaction of ion with the bio-molecular environment in a measurable way across a range of
intensity values of MF B. MFs do not have an appreciable effect on the rates of enzyme-catalyzed
reactions. Enzyme activity increases considerably when the MF is on, whereas activity decreases as
soon as the magnet is turned off. The activating effect is due to increase in hydrogen bonding and
consequently the helicity of the polypeptide backbone of the enzyme which stabilizes the protein
against denaturation; thus, the enzyme is less susceptible to inactivation (Akoyunglou, 1964).
The biggest requirement of food successfully preserved with MF technology is high electric resis-
tivity greater than 10–25 ohms-cm. Fortunately, many foods have electrical resistivity in this range.
The food systems preserved with MFs were: (a) milk with Streptococcus thermophilus, (b) yoghurt
with Sacchromyces, (c) orange juice with Sacchromyces, and (d) bread roll dough with bacterial
spores (Hofmann, 1985). The potential application of MF includes decontamination and recycling
of water on space flights tested by Chizhov et al. (1975).
14.3.2.1.1 Mechanisms of Microbial Inactivation
Hofmann (1985) used an OMF to produce PMF and found that the PMF was very effective in
deactivation of microorganisms in foods. In the study, the food was kept in a sealed plastic bag
and treated for various times from 25 to 100 ms at temperatures of 0°C–50°C using 1–100 pulses
of PMF at a frequency between 5 and 500 kHz. However, it has also been reported that the PMF
treatment had no significant effect on microbial populations (Malko et al., 1994; Caubet, 1999;
San Martı ́n et al., 2001) and even enhanced the microbial growth (Okuno et al., 1993). Also, static
PMF treatment was typically used in most of the study because, the MF at the sterilization cham-
ber inside the coil was not uniform, uniform sterilization in the test products may not be achieved.
However, if the product is under dynamic condition, the microorganisms in the product may have
increased opportunity to be uniformly exposed to MF and improve inactivation efficiency.
The effect of PMF intensity and pulse number on the bactericidal property of PMF in steriliza-
tion of fresh watermelon juice was studied by Ma et al. (2003b). The results showed that the overall
bactericidal effect was strengthened as the MF intensity and pulse number increased with the best
effect observed when the MF intensity was 2.53 T and pulse number was 20. Investigations spon-
sored by Bio-Magnetics Systems, Inc. have shown that unidirectional MFs inhibited or increased
the growth of cancer cells, depending on the field polarity. So, the effects of MFs were studied in
different areas such as drug delivery, cancer therapy, sterilization, and water treatment. MF affects
DNA synthesis and transcription as well as ion transcription through all membranes. The exposure
of the bacteria Serratia marcescens to a SMF 80 + 20 Gauss resulted in inhibition of growth. The
effect of MF was variable depending on the type of the microorganism and field. Study clarified that
MF has significant effect on bacterial cells as well as on its life and the effect of MF was enclosed
in the cell membrane. Other study found that E. coli, bacteria decarboxylation, and Staphylococcus
aureus viability affected the MF (10 mT, 50 Hz). The MF effect on bacteria could be considered as
bactericidal. Many researchers studied the effect of MF on bacteria (Kamel et al., 2014).
The MF changes the sensitivity of Pseudomonas aeruginosa to antibiotics and this agrees with
a study done by Ceon and Martin (2005), who found that exposure of E. coli to SMF increased its
antibiotic resistance. The biological effects of MFs may critically depend on the physical character-
istics of the magnetic signal, in particular the wave forces. So, treating enzymes with different MFs
can inhibit or promote enzyme activity according to API 20 E (Kamel et al., 2013). The growth rate
of Pseudomonas aeruginosa cells was affected by exposure to magnetic forces (400, 800, 1200, and
1600). In addition to that the MF increased the logarithmic phase within 4–6 h of treatment, but it
was decreased after 16–18 h as compared to the control. The bacterial enzymes such as TDA, GLU,
and ARA are affected by MF. So, it appears that treating enzymes with different MFs forces could
inhibit or promote enzyme activity according to API tests. Furthermore, the bacterial sensitivity to
antibiotics was increased after exposure period of 6 h to certain antibiotics, but became resistant
after 16 h (Kamel et al., 2013).
The importance of studying the effects of the MF upon the biological organisms was concret-
ized by the appearance of a scientific branch - the bio-magnetism. The MFs are of two types:
static (SMF) and oscillating (OMF). In case of SMFs, the intensity is constant in time, and in
case of OMFs, the intensity is varied, the constant amplitudes alternating with sinusoidal ampli-
tudes. Yoshimura (1989) classified the effects of MFs on microbial growth and reproduction as
inhibitory, stimulatory, and nonobservable. There are some research papers regarding the influ-
ence of the MF on the metabolism of the cellulolytic microorganisms. At the international level,
the research regarding the influence of the MF on the microorganisms have been conducted since
1937, when G.C. Kimball (1938) showed that the wine yeast cells were not affected by exposure for
5, 10, 20, 40, or 80 minutes to the MF. Gerencser et al. (1962) showed that the growth rate of the
Staphylococcus aureus species increases further to the exposure to a MF, for 3–6 hours, but the
exposure for 7 hours does not have any effect; the growth rate of the Serratia marcescens species
remains the same further to a 6 hours exposure to the MF, but it increases following an exposure
longer than 6 hours. Van Nostran et al. (1967) studied the effects of high MFs on the multiplication
of Saccharomyces cerevisiae, and Moore (1979) showed that the influence of the SMF has stimu-
lating effects on the growth of Escherichia coli, remarking that the growth stimulation is directly
proportional to the increase of the MF frequency. Tsuchiya et al. (1996) studied the effect of the MF
on the growth and development of Escherichia coli; also, Ruzic et al. (1997) performed exhaustive
researches regarding the influence of the MF on Pisolithus tinctorius.
Experiments have shown that strong static (SMF) or oscillating (OMF) MFs (5–50 T) have the
potential to inactivate vegetative micro-organisms. The impulse duration is between 10 ms and
several milli seconds. The frequencies are maximally 500 MHz, because above that value the items
begin to warm up noticeably. The effects of MFs on microbial populations have produced con-
troversial results (Tsuchiya et al., 1996). Before considering this technology for food preservation
purposes consistent results concerning the efficacy of the method are needed (Butz and Tauscher,
2002). Pothakamury et al. (1993) summarized the effect of MFs on microorganisms as shown in
Table 14.2.
table 14.2 Effect of Magnetic Fields on Microorganisms

Field Frequency
Micro- type of Strength of pulse
SlNo Organism Magnetic Field (t) (hz) Effect references
1. Wine yeast cell Heterogeneous 0.04 0 Growth inhibited Kimball
sMF when exposed for (1937)
5, 20, 25, 60,
120, or 150 min;
no inhibition for
10, 15, 17 min
exposure
2. Wine yeast cell Heterogeneous 1.1 0 no effect for 5, 10, Kimball
sMF 20, 40, or 80 min (1937)
exposure
3. Serratia Heterogeneous 1.5 — Growth rate Gerencser
marcescens sMF remains same as et al. (1962)
in controls up to
6 h; growth rate
decreases
between 6 and
7 h and again
increases
between 8 and
10 h; at 10 h cell
population same
as in controls
4 Staphylococcus Heterogeneous 1.5 0 Growth rate Gerencser
aureus sMF increases between et al. (1962)
3 and 6 h; then
decreases
between 6 and
7 h; cell population
at 7 h is same as
controls
5 Saccharomyces Heterogeneous 0.465 0 rate of Van nostran
cerevisiae sMF reproduction et al. (1967)
reduced,
incubated for 24,
48, or 72 h
6 Escherichia coli sMF 0.3 0 Growth simulated Moore
(1979)
7 Halobacterium sMF 0.015 0 Growth inhibited Moore
halobium, 0.03 (1979)
Bacillus 0.06
subtilis
8 Pseudomonas oMF 0.015 0.1–0.3 Growth simulated; Moore
aeruginosa, 0.03 stimulation (1979)
Candida 0.06 increases with
albicans increase in
frequency
9 E. coli oMF 0.15 0.05 inactivation of cells Moore
when (1979)
concentration
was 100 cells/mL
10 Streptococcus oMF 12.0 6,000 Cell population Moore
themophilus in (1 pulse) reduced from (1979)
milk 25,000 cells/mL
to 970
(Continued)
table 14.2 (Continued) Effect of Magnetic Fields on Microorganisms
Field Frequency
Micro- type of Strength of pulse
SlNo Organism Magnetic Field (t) (hz) Effect references
11 Saccharomyces oMF 40.0 416,000 Cell population Hofmann
in yogurt (10 pulses) reduced from (1985)
3,500 cells/mL to
25
12 Saccharomyces oMF 40.0 416,000 Cell population Hofmann
in orange juice (1 pulse) reduced from (1985)
25,000 cells/mL
to 6
13 Mould spores oMF 7.5 8,500 Population Hofmann
(1 pulse) reduced from (1985)
3,000 spores/mL
to 1
14 Saccharomyces sMF 0.56 0 Decreased growth Van nostran
cerevisiae rate; interaction et al. (1967)
between
temperature and
MF only during
the logarithmic
phase
Source: Pothakamury, u.r. et al., Food Technol. 47, 85–93, 1993.
According to Hofmann (1985), only one pulse of OMF was adequate to reduce the bacterial
population between 10 and 10 cfu/g. The intensity of the MF required to achieve these effects varied
between 225 T and a frequency range from 5500 Hz. Inconsistent results have been obtained on the
effect of OMF on microbial growth (Table 14.2). In some cases, OMF stimulated or inhibited micro-
bial growth and, in others, it had no effect on microbial growth. The results presented in Table 14.2
show that, although not well understood, the effect of MFs on the microbial population of foods may
depend on the MF intensity, number of pulses, frequency, and property of the food (that is, resistiv-
ity, electrical conductivity, and thickness of the foodstuff).
SMF or OMF may have some potential to inactivate microorganisms in food. Pothakamury
et al. (1993) reported two theories to explain the inactivation mechanisms for cells placed in SMF or
OMF. The first theory stated that a “weak” OMF could loosen the bonds between ions and proteins.
Many proteins vital to the cell metabolism contain ions. In the presence of a steady background MF
such as that of the earth, the biological effects of OMF are more pronounced around particular fre-
quencies, the cyclotron resonance frequency of ions (Coughlan and Hall, 1990). The second theory
discussed the effect of MFs on calcium ions bound in calcium-binding proteins such as calmodulin.
The Ca2ϩ ions vibrate regularly to an equilibrium position in the binding site of calmodulin. A steady
MF applied to calmodulin results in the rotation of the plane of vibration, or proceeds in the direc-
tion of MF at a frequency that is exactly similar to the cyclotron frequency of the bound calcium.
Adding a “wobbling” MF at the cyclotron frequency disturbs the precision to such an extent that it
loosens the bond between the calcium ion and the calmodulin. The effect of MFs on the microbial
population of foods may depend on the MF intensity, number of pulses, frequency, and property of
the food (i.e., resistivity, electrical conductivity, and thickness of the foodstuff).
Frequencies higher than 500 kHz are less effective for microbial inactivation and tend to heat
the food material (Barbosa Cánovas et al., 1998). MF treatments are carried out at atmospheric
pressure and at moderate temperatures. The temperature of the food increases 25°C. According to
Hofmann (1985), exposure to MFs causes inhibition in the growth and reproduction of microorgan-
isms. OMF of intensity of 5–50 Tesla (T) and frequency of 5–500 kHz was applied and reduced
the number of microorganisms by at least 2 log cycles. Within the MF of 550 T, the amount of
energy per oscillation coupled to 1 dipole in the DNA is 10–10 eV (Hofmann, 1985). OMF of this
intensity can be generated using: (1) super conducting coils, (2) coils which produce DC fields, or
(3) coils energized by the discharge of energy stored in a capacitor (Gersdorf et al., 1983). Inhibition
or stimulation of the growth of microorganisms exposed to MFs may be a result of the MFs them-
selves or the induced electric fields. The latter is measured in terms of induced electric field strength
and induced current density. To differentiate between electric field and MF effects, a cylindrical
enclosure containing cells and a medium that can be adapted to in vitro studies employing uniform,
single phase, or extremely low frequency (ELF) MFs is recommended.
14.3.2.2 Pasteurization
The basis of food preservation lies in inactivation of enzymes and microorganisms. The micro-
bial growth can either be stimulated or ceased during exposure to MFs. Retardation of growth and
reproduction of microorganisms might be due to change in deoxyribose nucleic acid (DNA) synthe-
sis, a change in the orientation of bio-molecules and bio-membranes to a direction parallel or perpen-
dicular to the applied MF, or a change in the ionic drift across the plasma membrane (Pothakamury
et al., 1993). The first approach of food preservation by OMF was initiated by Hofmann (1985) with
a US patent. The basic requirement for the application of OMF technology for food preservation is
that the food should have high electrical resistivity, greater than 10–25 ohms-cm.
Many foods have electric resistivity in these ranges. One most common example of food that can
be processed by OMF technique is orange juice. The electrical resistivity of orange juice is about
30 ohms-cm. The field intensity of the food to be magnetized is a function of electrical resistivity
and sample thickness. However, no correlation has been observed between MF intensities and food
constituents. Inactivation of microorganisms in food has been reported to occur during exposure
to OMFs with intensity higher than 2 T (Hofmann, 1985). A single pulse intensity of 5–50 T and
frequency of 5–500 kHz reduced the initial number of microorganisms by 2 log cycles. The technol-
ogy could be applied for food pasteurization purposes by placing the sample in MF and magnetized.
14.3.2.3 Sterilization
PMF sterilization is a new kind of physical cold sterilization technology, which has the char-
acteristics of low temperature rise of bactericidal material, low power consumption, no magnetic
leakage, high efficiency, high sterilization efficiency, easy control, strong penetrating ability, and
thorough sterilization. This technology is now widely used in the circulating water sterilization of
air conditioning refrigeration systems. The PMF sterilization has a broad market potential in the
cold sterilization technology because the method does not need heat and costs less time. A large
number of studies have indicated that high-strength PMF sterilization has important application
value in liquid food sterilization industry. But in the current PMF device of air conditioning system
circulating water, the applied discharge voltage is too high, generally from thousands of volts to
tens of thousands of volts. Such a high voltage has brought great inconvenience to the operator and
safety performance is also relatively poor; therefore, these problems have hindered the research and
development of PMFs in the field of sterilization and limited their application and promotion in the
food industry.
The PMF technology showed that sterilization method can effectively kill the bacteria and fungi
in the fruit juice under the premise of retaining the nutritional components of fruit juice compared
with high-temperature sterilization. The effect of sterilization is proportional to the time of steril-
ization, and the sterilization effect of bacteria is better than that of the fungus. The research on the
low-voltage electromagnetic pulse sterilization method would promote the nonthermal sterilization
of the PMF fruit juice to the industrial application. Sterilization effects of the PMF with a maxi-
mum intensity of 11.37 T were investigated on Escherichia coli AS 1.129, Staphylococcus aureus
AS 1.89, Saccharomyces cerevisiae ATTC 7552, and Bacillus subtilis AS 1.921. The well-regulated
fluctuations of sterilization effects with MF intensity and pulse number were observed, and can
be described by the “window effect” of MFs and provide a better explanation of the inconsistent
results of PMF sterilization. Sensibility of bacteria on the PMF significantly depends on a variety
of microorganisms. Sterilization effects of a flowing sample were better than that of static samples
(Ma et al., 2008).
Guozhang and Xiaohui (1998) reported that the nonthermal biological effect of MFs with low
intensity and frequency (Hz) could impact the sterilization results. Therefore, it is important to
determine the effect of PMF parameters under static and dynamic conditions on sterilization of
various microbes. The studied PMF parameters included MF factors, such as intensity and pulses
(Yang et al., 2004); product properties, such as ionic concentration, temperature, and pH (Ma et al.,
2004); and physiological factors, such as microbial cell concentration. The sterilization effect
of PMF treatment on bacteria at different population growth stages and the change and distri-
bution in temperature of model food liquid containing bacteria in the sterilization chamber were
also measured (Ma et al., 2004). The treated materials included model liquid containing E. coli,
S. aureus, S. cerevisiae, B. subtilis (Cao et al., 2003; Ma et al., 2004), milk (Gao et al., 2005), fore-
milk (Luo and Ma, 2004), watermelon juice (Gao et al., 2004), and beer (Ma et al., 2003a). These
studies provided fundamental information about the effectiveness of PMF treatment for steriliza-
tion, which states that the treated materials and intensity of PMF could be important factors in
using the nonthermal treatment method (Ma et al., 2008). This study verified the sterilization effect
of the low-pressure PMF on the fruit juice products. According to the sterilization effect test of
low-pressure PMF equipment on fruit juice, the results showed that the low-pressure PMF possessed
a bactericidal effect on fruit juice, and the sterilization effect improved with increase in time.
Gerencser et al. (1962) reported on the inactivation effect of MFs on bacteria. Chizhov et al. (1975)
discovered 80% decrease of total numbers of Escherichia coli in water after MF treatment at power of
0.5–1.5 Tesla (T). Hofmann (1985) reported deactivation of microorganisms by OMF. Pothakamury
et al. (1993) found inactivation effect of OMFs on packed food at the pulse number of 1–100, tempera-
ture of 0°C–50°C, and time of 20–100 ms. Mehedintu and Berg (1997) reported 16% inactivation and
25% of yeast Saccharomyces cerevisiae by 0.2 and 0.5 mT MF, respectively. Ma et al. (2003a, 2003b)
discovered the inactivation effect of microorganisms in crude beer and watermelon juice by PMF
sterilization. However, San Martı ́n et al. (2001) and Harte et al. (2001) did not find any inactivation
effect of high-intensity MF on E. coli ATCC 11775 even with 18T high power. Little attention has been
paid to these contrary results on PMF sterilization, and no reasonable interpretation has been reported
about this phenomenon. Niu et al. (2003) reported that the biological window effect may be one of the
possible reasons based on their research. Biological window effect is one of the typical nonthermal
effects of ELF pulsed electromagnetic waves on the cell. Biological window effect includes the effects
of frequency window, intensity (power density) window, and other windows. The former refers to
the fact that biological effects on cells can only be generated by those electromagnetic waves which
are separated and in very narrow frequency range in a particular frequency band; the corresponding
frequencies are called “the window of the frequency”. The latter one refers to the fact that biological
effects of electromagnetic waves will only be generated as discrete and with very narrow range of
intensity; and the corresponding intensity (power density) is the window of intensity. The presence of
these biological window effects has been proved by many tests (Niu et al., 2003). Bawin et al. (1978)
made an electromagnetic radiation on new chicken brain cells with a modulated very low-intensity
low-frequency of microwave, and found that biological effect, the release of calcium ions from the
cells, responds to a “window” effect of electromagnetic field intensity and frequency; that is, the
biological effect only responds to the microwave of 0.8 mW/cm2 and in the vicinity of 16 Hz. There
will be no response beyond this scope. They call the former (0.8 mW/cm2) the “window of strength”
and the latter (16 Hz) the “window of frequency”. Blankman et al. (1998) reproduced this experiment
and found that the windows of frequency are a series (i.e., more than one). Byus (1984) also found the
“window of time.” The contrary results of PMF sterilization with different powers may be caused by
the biological window effect of intensity, i.e., the inactivation effects on cells can only be generated
by those electromagnetic waves which are separated and in very narrow frequency range in a particu-
lar frequency band. However, no experimental result has been reported about the biological window
effect on cell in high-intensity band electromagnetic wave.
Self-designed PMF sterilization equipment Figure 14.2 (Chinese patent No. ZL 03220551.1,
whose power is up to 4.5T) was employed in this research. It consists of pulsed magnetic generator
and sterilization system which includes a spiral coil, an 8 cm diameter sterilization chamber, and a
thermostat (Figure 14.2). The PMF generator supplies PMF to the spiral coil at the set intensity and
pulse frequency (i.e., pulse number in one second). In this study, each prepared culture tube was put
on a glass hopper and set at the center of the sterilization chamber to subject PMF treatment. The
objective of this research was to investigate the effect of high-intensity PMF on the inactivation of
Escherichia coli 8099 and to explore whether the biological window effects exist in PMF steriliza-
tion process (Byus et al., 1984).
The principle of low-pressure PMF sterilization apparatus is shown in Figure 14.2, which mainly
includes two parts of the pulse electric field and the PMF. The pulse current originated from PMF
system generated high-intensity PMF according to the solenoid coil of MF system, which could kill
bacteria in the materials. Electromagnetic field sterilization is utilizing electromagnetic energy to
destroy or affect the structure of the organism to achieve the purpose of eliminating or inhibiting it.
PMF is a kind of electromagnetic field, and in the process of sterilization of material it would
produce a variety of electromagnetic effects, which mainly include induced current effect, Lorenz
force effect, oscillation effect, ionization effect, and so on, and these electromagnetic effects can
cause cell biological effects, resulting in an important influence to the sterilization process. When
the PMF is sterilized, the duration of each pulse is very short, generally for a few microseconds to
a few milliseconds, and several pulses can make the material sterilization rate reach 90% or more.
Instantaneous sterilization is carried out under normal temperature and pressure by PMF, and the
process consists of four main elements: the MF intensity of the coil, pulse number, pulse current
waveform characteristics, and material characteristics. The pulse electric field generating system
is the main part of the low-pressure PMF sterilization device, which mainly consists of charging
circuit part, discharge circuit part, and control part. This circuit chooses a linear solenoid in which
r = 30 mm, l = 200 mm. The experiment shows that the PMF produced by the device can achieve a
better effect in killing bacillus coli, beer yeast and staphylococcus aurous, etc. Compared with the
Figure 14.2 Principle diagram of low-pressure PMF sterilization test apparatus. (From byus, C.V. et al.,
Bioelectromagnetics, 5, 314–351, 1984.)
traditional sterilization device, the bactericidal effect of the PMF sterilization device is more obvi-
ous so that it will be the direction of the development of food equipment (Otero et al., 2016).
The sterilization effect of PMF improved with the sterilization time whatever voltage was
loaded. The comparison of three kinds of different voltage indicated that the higher the voltage
loaded, the higher the sterilization rate was. The reason was that the higher the voltage loaded, the
stronger the MF strength was. This strengthened the penetration of the microorganisms, resulting
in improvement of the sterilization efficiency.
The PMF had sterilization effect on the above bacteria, and the sterilization effect gradually
improved with the increase of sterilization time. It is worth noting that the sterilization effect of
PMF on bacteria such as lactobacillus and colibacillus was better than the effects on fungi such as
mildew and microzyme. This was the result of the tolerance difference between the bacteria and
the fungus. The cellulosic cell wall of fungi had a strong protective effect, but the toughness of cell
wall in bacteria is less than that of fungus, and thus the tolerant ability of bacteria was poorer than
that of fungi (Zhang, 2016).
14.3.2.4 Magnetic Freezing
Magnetic freezing is able to generate tiny ice crystals throughout the frozen product, prevent
cell destruction, and preserve the quality of fresh food intact after thawing. If all these advantages
were true, magnetic freezing would represent a significant advance in freezing technology, not only
for food preservation but also for cryopreservation of biological specimens, such as cells, tissues,
and organs. MFs are supposed to act directly on water by orientating, vibrating, and/or spinning
molecules to prevent them from clustering and, thus, promote supercooling. However, many doubts
exist about the real effects of MFs on freezing and the science behind the potential mechanisms
involved. Moreover, many of the comparisons between magnetic and conventional freezing are not
correctly designed to draw valid conclusions and wide ranges of MF intensities and frequencies are
unexplored. Therefore, more rigorous experimentation and further evidence are needed to confirm
or reject the efficacy of MFs in improving the quality of frozen products (Table 14.3).
14.3.2.5 Yeast under High-Gradient Magnetic Field
Superconducting technology has provided new methods for studying the application of strong
MFs in biological systems and diamagnetic or paramagnetic materials. Room temperature bores of
superconducting magnets have given opportunities to explore the importance of the diamagnetism
of biological materials as well as that of strong (ferro) magnetism in solid state materials. A new
approach using superconducting technology involved gradient MFs in the horizontal direction,
which made several interesting phenomena. It was reported that the surface of water bends down-
ward under the MF of 10 T or so at the center. The parting of water has been termed the “Moses
effect”, a phenomenon in which the surface of water has been separated under high MFs. The hori-
zontal gradient magnetic force and the vertical gravitational force generate a sloping gravity-like
field for living organisms, in which the sloping field is considered to be the actual direction of grav-
ity. In the case of the parting of water, the water was pushed to lower MFs from higher fields, and
localized in the middle part of the space forming a “water wall”. However, the applied field should
be high and steep enough to clearly observe these effects. The effects of gradient MFs on the behav-
ior of yeast (S. cerevisiae) proliferation and mass distribution have been reported also by Iwasaka
et al. (2004) using strong SMFs (flux density 14 T). When yeast was exposed to 9–14 T MFs with
a maximum flux density gradient of dB/dx σ 94 T/m, where x is the space coordinate, the rate of
yeast proliferation under the MFs decreased after 16 h of incubation compared to that of the control.
The results indicated that the gas pressure inside a flask with 6 T, 60 T/m gradually increased in
comparison to the pressure inside a control tube. Owing to the diamagnetism of medium water and
276
table 14.3 patents on Magnetic Freezing

Magnetic Field Combinations
Electro
pulsed B; ω; Electric Magnetic Freezing products to
Sl. No. authors Static B Oscillating B; ω pw Fields E; ω Waves ω Others temperature Freeze Status
1. Hirasawa ω/γ — — — 0.01– Air sanitizer −40 °C Food Patented

and 100 MHz
others
(2000)
2. Hirasawa X X — — — Air sanitizer −20°C Food Patented
and
others
(2001)
3. owada 0.1 mt–2 t 0.1–10 mt; — 100– — sound wave −30°C to Food and food Patented
and 50–60 Hz 1000 kV/m generator −100°C ingredients
Kurita 10 mt 0.5 mt; — 600 kV/m — Ventilators −50°C tuna, sardines, tested
(2001) 50–60 Hz Far-infrared- pork, juices,
ray wine, oranges,
absorber and cakes
Heat
insulators
4. Kino X X — — γ·b not X Food Patented
(2002) described
5. owada 0.1 mt–1 t 0.1–100 mt; — 10–500 kV/m; — ionic air −20°C to −40°C Foodstuffs, food Patented
(2007) 50–60 Hz 50 Hz–5 MHz Ventilators products,
(0.25–3 MHz) Honeycomb organisms, and
Far-infrared- other materials
1 mt 0.5–0.7 mt; — 15 kV/m; — ray −20°C, −40°C Chicken and tuna tested
50 Hz 250 kHz, absorber
3 MHz, Heat
50 Hz-5MHz insulators
6. toyoshima X — — — X Ventilators −40°C Food Patented
(2005) Heat
insulators
7. ino and — ≤100 mt; — — — Ventilators X Food Patented
others ≤10 MHz Heat
(2005) — X — — — insulators −35°C sweet potatoes tested
(Continued)
table 14.3 (Continued) patents on Magnetic Freezing

Electro
8. Miura and — — 0.1–10 t; not −10°C to −60°C starch-containing Patented
others 0.1 Hz–1 kHz; described food
(2005) 10–100 μs
5t −20°C Potato starch tested
1 Hz gels
55 μs
9. sato and — 10 mt–1.2 t — — — ionic air −20°C to −60°C Food Patented
Fujita (30–700 mt)r; ultrasonic
(2008) 20 Hz–25 kHz waves,
PuLseD MAGnetiC FieLD ProCessinG oF FooDs
(40 Hz–1.2 kHz)r microwaves,

10 mt–1.2 t 10 mt–1.2 t — — — far infrared −20°C to −60°C Food Patented
(30– (30–700 mt)r; rays,
700 mt)r 20 Hz–25 kHz ultraviolet
(40 Hz–1.2 kHz)r light,
800 mt 300 mt; — — — α-rays −40°C, −50°C Chinese noodles, tested
100 Hz Air spinaches,
dehumidifier packed pasta,
Air pressure pork lumps, and
regulator tofu blocks
Ventilators
10 Kim and — X — — — not X ice cubes Patented
others described
(2009)
(Continued)
277
table 14.3 (Continued) patents on Magnetic Freezing
278

Electro
11 owada 0.1 mt–2 t 0.1–10 mt — 100– — Air pressure −30° to −100 °C Food products, Patented
and saito (10 mt)r (0.5 mt)r; 1000 kV/m regulator food ingredients,
(2010) 50–60 Hz (2–60 kV/m)r Air sanitizer medical
oxygen products,
absorber medicines, living
sound wave tissues, and living
generator cells
X X — X — Ventilators X Mackerel and tested
Far-infrared- lobster
ray
absorber
Heat
insulators
12 Fujisaki 1–200 mt — — — 0.3–2 MHz Ventilators X Food products, Patented
and (10– (0.6–1 MHz) Far-infrared- cooking
Amano 15 mt)r ray ingredients, living
(2012) absorber bodies, and
Heat biological
insulators samples
13 Mihara — 0.01–0.4 mt — — — not −2°C to −40°C Foods, organs, Patented
and (0.2 mt)r; described (−20°C to and the like
others 200 Hz–200 kHz −40°C)r
(2012) (2 kHz)r
0.12 mt; — — — X Physiological tested
0 Hz–200 kHz saline solution
0.1–0.2 mt; — — — −30°C rats
2 kHz
0.8 mt; — — — −40°C Alkaline
2 kHz phosphatase,
green
fluorescent
protein
Source: otero, L. et al., Compre. rev. Food sci. F., 15, 646–667, 2016.
Note: b: MF strength; ω: Frequency; pw: Pulse width; e: electric field strength; γ: gyromagnetic ratio for hydrogen (42.58 MHz/t); —: not employed; X: not reported value;
()r: recommended conditions.
yeast, the liquid surface clearly inclined under gradient MFs, and the hydrostatic force in suspen-
sion was strengthened by the diamagnetic forces. In addition, magnetophoresis of the yeast cells in
the medium solution exhibited localization of the yeast sedimentation pattern. The mechanisms for
deceleration of yeast proliferation by MFs have been proposed and well described by Iwasaka et al.
(2004) and Ahmed and Ramaswamy (2007).
14.3.2.6 Isolation and Separation of Protein by Magnetic Technique
Isolation and separation of specific molecules is common practice in almost all areas of bio-
sciences and biotechnology. Various techniques have been used to achieve this goal. Recently,
increased attention has been paid to the development and application of magnetic separation tech-
niques, which employ small magnetic particles. Safarik and Safarikova (2004) recently reviewed
magnetic techniques for the isolation and purification of proteins and peptides. The basic prin-
ciple of batch magnetic separation is simple. Magnetic carriers bearing an immobilized affinity or
hydrophobic ligand or ion-exchange groups, or magnetic biopolymer particles having affinity to the
isolated structure, are mixed with a sample-containing target. Magnetic separation techniques have
several advantages over standard separation techniques. This process requires only a few handling
steps. The separation can be carried out directly in crude samples containing suspended solid mate-
rial. Owing to the magnetic properties of magnetic adsorbents (and diamagnetism of most of the
contaminating molecules and particles), they can be relatively easily and selectively removed from
the sample. In fact, magnetic separation is the only feasible method for recovery of small mag-
netic particles (diameter of 0.1–1 m) in the presence of biological debris and other fouling material
of similar size. In addition, the power and efficiency of magnetic separation procedures are use-
ful in large-scale operations. Several automated systems for the separation of proteins or nucleic
acids have become available recently. Magnetic separation is usually a very mild operation to the
target proteins or peptides. Even large protein complexes may remain intact when using the very
gentle magnetic separation procedure. The separation process has been significantly influenced
by both the reduced shearing forces and the higher protein concentration throughout the isolation
process. Appropriate magnetic particles can be used for their concentration instead of ultrafiltration,
precipitation, etc (Ahmed and Ramaswamy, 2007).
14.3.2.7 Magnetic Resonance Imaging
The applications of MRI technology (magnetic resonance used) are growing very fast and
could be used for food quality control purposes in the future. MRI is an imaging technique used
basically in health care technology to produce high-quality images of the inside of the human
body. MRI is based on the principles of NMR, a spectroscopic technique used by scientists to
obtain microscopic chemical and physical information about molecules. However, the tremen-
dous potential of magnetic resonance systems in other fields has not been fully explored yet. The
technique was called MRI rather than nuclear magnetic resonance imaging (NMRI) (Ahmed
and Ramaswamy, 2007).
14.3.3 application of Magnetic Fields in Quality Control of Food products
• MRI is an attractive tool for temperature mapping (distributions) induced in water-based foods
by microwave and conductive heating. Also, it can measure quantitatively in three dimensions all
aspects of the mass transport of water and fat in foods to give direct information about the effects of
process engineering, including flow, mixing, and heating.
• MRI has been proven efficient as an inline sensor for detecting defects and measuring the quality
of fruits and vegetables.
• MRI can predict the structure and dynamics of foods and other heterogeneous materials during and
after processing.
• MRI has been used efficiently as a viscometer; MRI measurements of the fluid velocity profile in tube
flow are coupled with a pressure drop measurement to yield shear viscosity. Each measurement yields
a range of shear viscosity data for a single flow rate, as compared to most conventional viscometers
that produce only one data point under the same circumstances. This technique has the potential to
significantly enhance process control of industrial processes (Ahmed and Ramaswamy, 2007).
14.4 CONCLUSIONS aND FUtUrE trENDS
Nonthermal processing is the latest application in food technology and in the twenty-first cen-
tury most of these technologies, which were restricted to laboratory studies in the past, are being
commercialized. These nonthermal technologies can revolutionize food processing as such and in
the improvement of batch capacities and the development of continuous systems could be highly
essential to realize the full commercial potential of these highly advanced techniques and to reduce
high-cost capitalization and also to make the product cost effective. Food engineering has a chal-
lenging task ahead in making these technologies and equipment available for introduction in the
developing countries. The PMF technology can not only result in restriction in microbial loads but
also could be extremely useful as a process aid in terms of protein conformational changes and
overall improvement in sensory and nutritional attributes. However, to meet this high expectation,
consumers and stakeholders must be convinced about the improvements in these new technolo-
gies. This will require convincing data and provision of clear, objective, and unbiased information
including the potentially negative aspects of the technology and their limitations. The use of MFs as
an alternative food processing technology has not gained full commercial acceptance possibly due
to minimally processed foods, and therefore food processing sectors are keenly looking for incon-
sistent results on microbial growth and death kinetics. In addition, there is a significant lack of infor-
mation on the field and the development of its machinery. Application of MFs could be effective and
beneficial for biomass production at controlled growth rate using superconducting technology. This
would help the growth of fermentation and pharmaceutical industries. The structural modifications
of proteins and fats could be achieved by exposing these constituents in MFs.
rEFErENCES
Ahmed, J., and Ramaswamy H.S. (2007). Applications of magnetic field in food preservation. In: Handbook
of Food Preservation, pp. 855–866. Rahman, M.S. Ed., CRC Press, New York.
Akoyunglou. (1964). Effect of a magnetic field on carboxydismutase. Nature. 202: 452–454.
Barbosa-Cánovas, G.V., Gongora-Nieto, M.M., and Swanson, B.G. (1998). Non-thermal electrical methods in
food preservation. Food Sci Technol Int. 4: 363–370.
Barbosa Canovas, G.V., San Martin, M.F., Harte, F., and Swanson, B.G. (2001). Magnetic fields as a
potential non-thermal technology for the inactivation of microorganisms. In: Control of Foodborne
Microorganisms, pp. 399–418. Juneja, V.K., and Sofos, J.N. Eds., Marcel Dekker, New York.
Bawin, S.W., Adey, W.R., and Sabbot, I.M. (1978). Ionic factors in release of 45 Ca2+ from chicken cerebral
tissue by electromagnetic fields. P Natl Acad Sci USA. 75: 6314–6318.
Blankman, C.F., Benance, S.G., Elliott, D.J., House D.E., and Pollock, M.M. (1998). Influence from
brain tissue in Vitro: A three-model analysis consistent with the frequency response up to 510 Hz.
Bioelectromagnetics. 9: 215–227.
Busta, F.F., Datta, A.K., Kokini, J.L., Pflug, I.J., and Pierson, M.D. (2000). Kinetics of Microbial Inactivation for
Alternative Food Processing Technologies. Research Report of U. S. Food and Drug Administration, June 2.
Butz, P., and Tauscher, P. (2002). Emerging technologies: Chemical aspects. Food Res Int. 35: 279–284.
Byus, C.V., Lundak, R.L., Fletcher, R.M., and Adey, W.R. (1984). Alterations in protein kinase activity fol-
lowing exposure of cultured human lymphocytes to modulated microwave field. Bioelectromagnetics.
5: 314–351.
Cao, H., Ma, H., Cui, H., and Liu, T. (2003). Sterilization effect of the pulsed magnetic field on Escherichia
coli and Staphylococcus aureus. Sci Technol Food Ind Chinese. 4: 30–33.
Caubet, R. (1999). Effets des champs magnetiques pulses an haute densite de flux sur les micro-organismes.
In: Proceedings of the Technological Training in the Food Industry, Conservation for Tomorrow,
2nd ed. Bordeaux Pessac, France.
Ceon, R., and Martin, J.T. (2005). Low-level, magnetic field induced growth modification of Bacillus subtilis.
J Electromagnet. 8: 275–82.
Chizhov, S.V., Siniak, I.E., Shikina, M.I., Ukhanova, S.I., and Krasnoshchekov, V.V. (1975). Effect of a mag-
netic field on Escherichia coli. Kosm Biol Aviakosm Med. 9: 26–31.
Coughlan, A., and Hall, N. (1990). How magnetic field can influence your ions? New Sci. 8: 30.
Date, M. (1983). High Field Magnetism. Proceedings of the International Symposium on High Field
Magnetism, Osaka, Japan, September 13–14, North Holland, Amsterdam, the Netherlands.
Davis, A.R., and Rawls, J.R. (1974). Magnetism and its Effects on the Living System, Acres, U.S.A., Kansas
City, MI.
Farkas, J. (2007). Physical Methods of food preservation. In: Food Microbiology: Fundamentals and Frontiers, pp.
685–712. Doyle, M., and Beuchat, L. Eds., ASM Press, Washington, DC. doi:10.1128/9781555815912.ch32.
Gao, M.X., Ma, H.L., and Guo, K.Q. (2004). Sterilization of watermelon juice using pulsed magnetic field.
Food Ferment Ind Chinese. 3: 14–17.
Gao, M.X., Ma, H.L., and Guo, K.Q. (2005). Sterilization effect of pulsed magnetic field and its influence on
milk nutrient components. Trans Chinese Soc Agr Eng. 21: 181–184.
Gerencser, V.E., Barnothy, M.F., and Bamothy, J.M., (1962). Inhibition of bacterial growth by magnetic fields.
Nature. 196: 539–541.
Gersdorf, R., deBoer, F.R., Wolfrat, J.C., Muller, F.A., and Roeland, L.W. (1983). The High Magnetic Facility
of the University of Amsterdam, High Field Magnetism. Proceedings International Symposium on High
Field Magnetism, Osaka, Japan, pp. 277–287.
Guozhang, C., and Xiaohui, C. (1998). Talk on research hot point of bio-electromagnetics: Non-thermal effect.
Phy Chinese. 27: 151–155.
Harte, F., San Martin, M.S., Lacerda, A.H., Lelieveld, H.L.M., Swanson, B.G., and Barbosa Canovas, G.V.
(2001). Potential use of 18 tesla static and pulsed magnetic fields on Escherichia coli and Saccharomyces
cerevisiae. J Food Process Pres. 25: 223–235.
Hofmann, G.A. (1985). Deactivation of microorganisms by an oscillating magnetic field. U.S. Patent 4,524,079.
http://www.fda.gov/Food/FoodScienceResearch/SafePracticesforFoodProcesses/ucm103131.htm 1/8,
http://www.paper.edu.cn.
Iwasaka, M., Ikehatab, M., Miyakoshic, J., and Uenoa, S. (2004). Strong static magnetic field effects on yeast
proliferation and distribution. Bioelectrochemistry. 65: 59–68.
Kamel, F.H., Saeed, C.H., and Qader, S.S. (2013). The effects of magnetic fields on some biological activities
of Pseudomonas Aeruginosa. Diyala J Med. 5: 29–35.
Kamel, F.H., Saeed, C.H., and Qader, S.S. (2014). Magnetic field effect on growth and antibiotic susceptibility
of Staphylococcus aureus. J Al-Nahrain Univer. 17: 138–143.
Kimball, G.C. (1938). The growth of yeast in a magnetic field. J Bacteriol. 35: 109–122.
Liboff, A., Williams, T., Strong, D.M., and Wister, R. (1985). Magnetic correlates in electromagnetic con-
sciousness. Electromag Biol Med Sci. 35: 228–236.
Luo, X., and Ma, H. (2004). Sterilization of cow foremilk using pulsed magnetic field. China Dairy Ind
Chinese. 32: 22–23.
Ma, H.L., Deng, Y.L., and Chu, J.Y. (2003a). Sterilization of crude beer with magnetic inspired pulse electro-
magnetic field and analysis of its mechanism. Chinese J Food Sci. 24: 52–55.
Ma, H.L., Deng, Y.L., and Chu, J.Y. (2003b). Sterilization of watermelon juice with high voltage pulse mag-
netic field and its mechanism analysis. Trans Chinese Soc Agr Eng. 19: 163–166.
Ma, H., Gao, M., and Guo, K. (2004a). Influence of medium parameters of microorganisms on bactericidal
effect of pulsed magnetic fields. Food Sci Chinese. 25: 42–46.
Ma, H., Pan, Z., Gao, M., and Luo, L. (2008). Efficacy in microbial sterilization of pulsed magnetic field treatment.
Int J Food Eng. 4(4): 1556–3758. doi:10.2202/1556-3758.1177.
Ma, H., Wu, Q., Gao, M., and Chu, J. (2004b). Influence of growth stage and medium parameters of micro-
organisms on bactericidal effect of pulsed magnetic fields. Trans Chinese Soc Agr Eng. 20: 215–217.
Malko, J.A., Constantinidis, I., Dillehay, D., and Fajman, W.A. (1994). Search for influence of 1.5 tesla mag-
netic field on growth of yeast cells. Bioelectromagnetics. 15: 495–501.
Martin, O., Vega-Mercada, H., Qin, B.L., Chang, F.J., Barbosa-Canovas, G.V., and Swanson, B.G. (1997).
Inactivation of E. coli suspended in liquid eggs using pulsed electric fields. J Food Process Pres. 21: 193.
Mathavi, V., Sujatha, G., Ramya, S.B., and Devi, B.K. (2013). New trends in food processing. Int J Adv Eng
Technol, IJAET. 5: 176–187.
Mehedintu, M., and Berg, H. (1997). Proliferation response of yeast Saccharomyces cerevisiae on electromag-
netic field parameters. Bioelectrochem Bioenerg. 43: 67–70.
Mertens, B., and Knorr, D. (1992). Developments of non-thermal processes for food preservation. Food
Technol. 46: 124–133.
Miura, N. (1991). Generation of megagauss magnetic fields and their application to solid state physics, phys-
ics and engineering applications of magnetism. In: Solid State Sciences, pp. 19–47. Ishikawa, Y., and
Miura, N. Eds., Springer-Verlag, Berlin, Germany.
Moore, R.L. (1979). Biological effects of magnetic fields. Studies with microorganisms. Can J Microbiol. 25:
1145–1151.
Niu, Z.Q., Hou, J.Q., Wang, H.B., Zhang, H., Yan, J., Lu, Z.Y., Guo, G.Z., Wang, C.M., and Lin, H. (2003). The
window bio-effects of electromagnetic waves. Chinese J Biol Eng. 22(2): 126–132.
Okuno, K., Tuchiya, K., Ano, T., and Shoda, M. (1993). Effect of super high magnetic field on the growth of
Escherichia coli under various medium compositions and temperatures. J Ferment Bioeng. 75: 103–106.
Otero, L., Rodríguez, A.C., Pérez-Mateos, M., and Sanz, P.D. (2016). Effects of magnetic fields on freezing:
Application to biological products. Compr Rev Food Sci F. 15: 646–667. doi:10.1111/1541-4337.12202.
Pothakamury, U.R., Barbosa Canovas, G.V., and Swanson, B.G. (1993). Magnetic field inactivation of micro-
organisms and generation of biological changes. Food Technol. 47: 85–93.
Rubin, L.G., Weggel, R.J., Leopold, M.J., Williams J.E.C., and Iwasa, Y., (1983). High field facilities at the
Francis Bitter National Magnet Laboratory. In: Proceedings of the International Symposium on High
Field Magnetism, Okasa, Japan, pp. 249–254. Date, M. Ed., North Holland, Amsterdam, the Netherlands.
Ruzic, R., Gogala, N., and Jerman, I. (1997). Sinusoidal magnetic fields: Effects on the growth and ergosterol
content in mycorrhizal fungi. Electro- Magnetobiol. 16: 129–142.
Safarik, I. and Safarikova, M. (2004). Magnetic techniques for the isolation and purification of proteins and
peptides. Biomagn Res Technol. 2(7): 1–17.
San Martı ́n, M.F., Harte, F.M., Lelieveld, H., Barbosa-Cánovas, G.V., and Swanson, B.G. (2001). Inactivation
effect of an 18-T pulsed magnetic field combined with other technologies on Escherichia coli. Innov
Serway, R.A. (1982). Physics: For Scientists and Engineers. CBS College Publishing, New York.
Sloan, A.E. (1999). Top ten trends to watch and work on the millennium. Food Technol. 53: 40–58.
Tsuchiya, K., Nakamura, K., Okuno, K., Ano, T., and Shoda, M., (1996). Effect of homogeneous and inhomo-
geneous high magnetic fields on the growth of Escherichia coli. J Ferment Bioeng. 81: 343–346.
Van Nostran, F.E., Reynolds, R.J., and Hedrick, H.G. (1967). Effects of a high magnetic field at different
osmotic pressures and temperatures on multiplication of Saccharomyces cerevisiae. Appl Microbiol.
15: 561–563.
Wouters, P.C., and Smelt, J.P.P.M. (1997). Inactivation of microorganisms with pulse electric fields: Potential
for food preservation. Food Biotechnol. 11: 193–229.
Yang, Q., Gao, M., and Ma, H. (2004). Sterilization effect of pulsed magnetic field and its influence on food
quality. J Microw Chinese. 20: 82–85.
Yoshimura, N. (1989). Application of magnetic action for sterilization of food. Shokukin Kihatsu. 24: 46–48.
Zhang, Z. (2016). Data analysis of sterilization effect of low pressure pulse magnetic field on fruit juice
products. In: International Conference on Computational Science and Engineering (ICCSE), pp. 14–17.
Atlantis Press Publisher.
ChaptEr 15
Use of plasma in Food processing
A. K. Pandey and O. P. Chauhan
CONtENtS
15.1 Introduction ........................................................................................................................284

15.2 Plasma ................................................................................................................................284
15.3 Plasma Classification.......................................................................................................... 286
15.3.1 Thermal Plasma.................................................................................................... 286
15.3.2 Non-thermal Plasma ............................................................................................. 287
15.3.2.1 Sources of Non-thermal Plasma.......................................................... 287
15.4 Plasma in Food Processing................................................................................................. 288
15.4.1 Microbial Inactivation .......................................................................................... 288
15.4.2 Enzyme Activity................................................................................................... 291
15.4.3 Starch Modification .............................................................................................. 291
15.4.4 Seed Germination................................................................................................. 292
15.4.5 Functional Properties of Food .............................................................................. 293
15.5 Application of Plasma in Food Processing Industries........................................................ 293
15.5.1 Application in Cereal Industry ............................................................................. 294
15.5.2 Seed Germination................................................................................................. 296
15.5.3 Herb/Spice Processing Industry ........................................................................... 296
15.5.4 Fruits and Vegetables Processing Industry .......................................................... 298
15.5.5 Milk Processing Industry ..................................................................................... 299
15.5.6 Meat and Poultry Processing Industry .................................................................300
15.5.7 Application in Food Packaging Industry ............................................................. 301
15.6 Factors Affecting the Plasma Efficiency in Food Processing ............................................ 303
15.6.1 Gas/Mixture of Gases ..........................................................................................304
15.6.2 Direct and Indirect Treatment ..............................................................................304
15.6.3 Other Factors ........................................................................................................304
15.7 Limitations of Plasma Processing ...................................................................................... 305
15.8 Future Prospects of Plasma Processing..............................................................................306
References ......................................................................................................................................306
283
15.1 INtrODUCtION
Growing demand of safe and minimally processed food poses challenges to the food
processing and supply system. Food safety is a major concern for regulatory agencies as well as
for food industries. Therefore, need of sustained quality characteristics of food without compro-
mising their sensory attributes, nutritional and functional properties, and assurance of enhanced
microbiological food safety has accelerated the interest of food processing industries in innova-
tive technologies. In the past, several methods were applied to eliminate microorganisms from
foods; these include lethal heat treatments such as sterilization, pasteurization, ohmic heating,
etc. There are multiple quality issues related to the inherent characteristics of food during ther-
mal processing such as reduction of nutrients, texture, colour, flavour, etc. This has led to the
emergence of non-thermal technologies (Sampedro et al., 2005). The negative effects of thermal
methods on nutritional and sensory characteristics of food can be minimized by the application
of non-thermal technologies which are efficient at sub-lethal or ambient temperatures (Tiwari
et al., 2009). Non-thermal technologies include application of high pressures (HP), pulsed elec-
tric field (PEF), pulsed light, irradiation, oscillating magnetic field (OMF), ultrasonics, ozona-
tion, plasma processing, etc. Most of these technologies require high initial investments, whereas
some are associated with adverse perceptions and/or other constraints, limiting the uses of these
commercially viable methods (Yun et al., 2010). The industries from various fields (e.g., textile,
electronics, life sciences, packaging, etc.) are moving towards plasma processing technology,
where products are treated with plasma to reduce the microbial load primarily from the surface,
leaving negligible or moderate impact on substrate material. Plasma is also being adopted com-
mercially as a replacement for disinfecting chemicals that are generally used for cold sterilization
of bio-medical devices and other thermo-labile materials manufactured from heat sensitive plas-
tics such as low-density polyethylene (LDPE), high-density polyethylene (HDPE), polypropylene
(PP), etc. (Soloshenko, 2000; Moisan et al., 2001). Moreover, former methods of sterilization
including chemical and thermal treatments are conventionally practiced for surface disinfection
of spices, seeds, fruits, and vegetables. These conventional methods are often time consuming
and leave some toxic residues on the surface. Plasma processing of food is recently recognized
as one of the emerging techniques for disinfection and modification of food properties. Based on
the properties, plasma (mainly local thermodynamic equilibrium [LTE] and non-local thermody-
namic equilibrium [non-LTE] plasma) is most recently added to the non-thermal food processing
category. The laboratory scale observation shows the possibilities of NTP application for decon-
tamination of fresh and processed foods. Plasma has been found more effective in sanitizing
the heat sensitive products including fresh milk, fruits and vegetables, meat and poultry, and
the surface of medical devices (Van de Veen et al., 2014). Plasma treatment is also investigated
for their effect on seed germination, inactivation of enzymes, physico-chemical modification of
food properties including modification of starch and protein, and in-pack sterilization of products
(Thirumdas et al., 2014).
15.2 pLaSMa
The term plasma can be defined as “a fourth state of matter next to the solid, liquid and gas”
(Figure 15.1). It is a partially or wholly ionized gas or a mixture of ionized gases that can be
produced by the exposure of a gas/gas mixture under different pressure (HP or low pressure or
atmospheric pressure) conditions using different electrical discharge sources. The highly reactive
gas species produced by the electrical discharge is called plasma that possesses electron, photon,
ions, free radicals, atoms, and metastables but have overall net neutral charges in its fundamental
or excited state.
use oF PLAsMA in FooD ProCessinG 285
Figure 15.1 stages of matter.
An American physicist, Irving Langmuir, first coined the term plasma in 1928 to narrate the
fourth state of matter which abundantly exists in the universe other than solid, liquid, and gas
(Langmuir, 1928). Increase in energy input of matter causes phase change from solid to liquid and to
gas; on the other hand, further increase in energy beyond a certain level of a gaseous state delivers an
excited stage of molecules, called plasma. It is a distinct state of a matter which refers to a partially or
completely ionized gas containing ions, free radicals, photons, and atoms in their parental or excited
states and possessing a net neutral charge. The net neutral charge of plasma is associated with positive
and negative charge carriers that are generally present in equal ratio (Kudra and Majumdar, 2009).
Plasma generation basically depends on dissociation of coupling of gaseous molecules where dis-
sociation energy varies in a wide range of temperature and pressure. This dissociation energy could
be mechanical, thermal, radiation, nuclear, or from the source of electrical current. The gaseous
molecules turn in to ions, electrons, free radicals, photons, neutrally charged atoms, and metastables
by the applied dissociation energy (Figure 15.2). These dissociated constituents of plasma are often
designated as “light” species, possessing essentially electrons and photons, in contrast of which the
Figure 15.2 Constituents of plasma.

rest of the constituents are designated as “heavy” species. The density and temperature of electrons
varies with the type and amount of energy supplied for the dissociation of gaseous molecules.
The first important element of plasma processing is ionization followed by factors like reaction
rate, rate constants, the mean free path, and the electron energy distribution (Fridman 2008). Based
on the reactions, the chemical process of plasma can be divided into two categories: homogenous
and heterogeneous gas phase reaction. For the processing of semiconductor materials particularly
heterogeneous chemical reactions are most important. In heterogeneous process, the chemical reac-
tion of the gas phase can be sub-divided in three categories: (i) etching or ablation, where material is
removed from the surface of the subject; (ii) chemical vapour deposition, in which material is added
on the solid surface during plasma polymerization; and (iii) physical or chemical modification, where
substrate surface is modified without addition or removal of material (D’Agostino et al., 2005).
15.3 pLaSMa CLaSSIFICatION
Plasma is categorized into two different classes on the basis of their generation as high-temperature
or fusion plasma and low-temperature plasma (Figure 15.3). The relative energetic state of atoms and
molecules such as electrons and heavy species are used for classification of plasma. In high-temperature
plasma all the species (ions, electrons, and neutral species) exist in a state of thermal equilibrium.
Low-temperature plasma is further sub-categorized into two different groups which vary on the basis
of their generation method: (i) thermal plasma (TP), which is also called quasi-equilibrium plasma;
and (ii) non-thermal plasma (NTP) or cold plasma (CP). These two groups of low-temperature plasma
are often present in a LTE and non-LTE state, respectively (Tendero et al., 2006).
15.3.1 thermal plasma
TP is characterized by its local thermal equilibrium state of ions, electrons, and neutral species
of gaseous molecules. It can be produced by means of direct or alternating current (DC or AC) or
Figure 15.3 Classification of plasma.

radio frequency (RF) or microwave source under HP (>10 kpa). It is frequently produced by the
devices known as plasma torches. In this type of plasma, electrons and the heavy species are pres-
ent in a typical thermodynamic equilibrium. TP is utilized as a spray for surface modification of
materials such as coatings of thick films or destruction of harmful-toxic substances (Bonizzoni and
Vassollo, 2002). It is also used for the treatment of industrial effluents, where material destruction
is hypothesized on two different concepts: (1) breakdown of chemical bonds by the application of
heat produced from plasma jet, (2) breakdown of chemical bonds through bombardment of highly
energized plasma particles on substrate molecules in combination with heat. Associated high work-
ing temperature and pressure makes TP unfeasible for the application in food processing as foods
are highly susceptible to heat.
15.3.2 Non-thermal plasma
The temperature of electrons much above that of the gaseous ions (microscopic temperature) is
characteristic of NTP. Based on temperature, energy level, and ionic density, NTP is also termed as
CP. It is generated by exposing a gas or a mixture of gases to an electric discharge under vacuum
or atmospheric pressure using DC or AC or RF or microwave as a power source. The generation
of atmospheric pressure NTP is typically illustrated as corona discharge, gliding arc discharge,
radio-frequency plasma discharge (RFP), and dielectric barrier discharge (DBD). Unlike TP, gen-
eration of NTP (near ambient temperature of 30°C–60°C) requires low power and pressure below
the atmospheric level (or under vacuum). In NTP, electrons and heavy species do not exist in ther-
mal equilibrium. Therefore, produced weekly ionized NTP has an important aspect and ability to
decontaminate object surface effectively without affecting their other characteristics. NTP can be
generated using atmospheric pressure plasma jet (APPJ), which has both technically and commer-
cially more economic interest of food industries as it requires ordinary conditions. The tempera-
ture of dissociated particles remains relatively cold in NTP because most of the electrical energy
channelled to electron component without heating the entire gas steam. This phenomenon makes
it suitable for different applications such as treatment of biological tissues and polymers where
high temperature is not desired (Nehra et al., 2008). In food processing industries, application of
energetic, reactive gases of CP on the food surface such as fruit, vegetable, meat, and poultry etc.
can effectively reduce microbial load (Banu, 2012).
15.3.2.1 Sources of Non-thermal Plasma
There are several methods developed for NTP discharge generation which vary on the basis of
their design and technical application in different sectors. Generally, NTP discharge can be used in:
(i) cleaning of environment, such as treatment of polluted air, water, and gas; (ii) biomedical fields,
such as for disinfection and sterilization of medical devices; and (iii) the field of food processing.
Initially, vacuum conditions were used for generation of NTP which was not feasible for industrial
operation due to high initial cost and other constraints. Later, NTP systems were improved to work
under atmospheric pressure that increased its industrial applicability due to its low cost and improved
treatment speed (Yoon and Ryu, 2007; Yun et al., 2010). The decontamination process became less
expensive and easier due to increased ability of generating atmospheric pressure NTP discharge under
ambient conditions (Kim et al., 2011). Most of the commercially available devices are designed for
research purpose mainly for biomedical applications. Therefore, convenient application of NTP in the
food sector requires NTP generating devices to be customized or tailor-made as per the requirement.
Some most frequently used NTP discharge techniques applicable to food industries are as follows:
DC glow discharge plasma: It is a simplest type of NTP source. In this method, plasma is gener-
ated by the application of operating gas between anode and cathode plates which are connected with
the DC electric source. This plasma process is used as a light source for processing of materials by
physical means such as vapour ion deposition, etching, and surface modification (Chiad et al., 2010;
Wissel et al., 2013; Faisal and Dawood, 2014; Chalkha et al., 2015).
RF discharge plasma: There are two types of RF discharge plasma categorized on the basis of
their coupling mechanism which includes capacitive and inductive coupled discharge. The plasma
is generated either by capacitive coupling or inductive coupling energy using AC power supply at
RF range (1 KHz–103 MHz; most commonly 13.56 MHz). In this type of system, temperature of
plasma ranges between 5000 K and 10,000 K whereas, pressure ranges typically between 1 and
10 Pa, whereas frequency range is typically between 1 and 100 MHz (19–21). This type of low-
temperature plasma discharge system is widely used for etching, deposition, and surface modifica-
tion for various heat sensitive materials in different fields including aerospace and microelectronics
(Rauf and Kushnerb, 1998; Lin and Adomaitis, 2001; Laimer and Stori, 2006; Bora et al., 2011).
Dielectric barrier discharge plasma: This type of system is based on a specific AC discharge
facility that generates plasma under atmospheric pressure at moderate gas temperature. DBD is
also known as barrier discharge or silent discharge. DBD plasma is generally utilized for various
important applications such as surface decontamination, sterilization, ozone generation, pollution
control, chemical vapour deposition, and surface activation etc. (Tanino et al., 2005; Kostov et al.,
2009; Shainsky et al., 2012; Cal and Schluep, 2014).
Surface discharge plasma: In this system pulsed operations or high voltage AC is applied to gener-
ate surface discharge. A typical surface discharge reactor consists of strip electrodes. These electrodes
are arranged in a series over the surface of alumina ceramic which is also embedded inside with a
thin metal electrode (which could be a planar or a cylindrical form). Generally, this technique is used
for surface modification and for the removal of air pollutants with high efficiency (Ruma et al., 2016).
Atmospheric pressure discharge plasma: In this, plasma discharge is generated using APPJ.
A mixture of gases like He, O2, Ar, etc. and others can be used as feed gas for plasma generation.
Atmospheric pressure plasma has wide area of application such as inactivation of microbes, ster-
ilization of medical tools, chemical vapour deposition, modification of polymers, and material
processing (Napartovich, 2001; Han et al., 2014; Hsu et al., 2015).
Microwave discharge plasma: This plasma discharge source is used for generating high-frequency
plasma in the range of electromagnetic radiations, i.e., GHz. It facilitates a wide range of opportunities
like low-temperature operation near the substrate level at a wide range of gas pressures, good unifor-
mity, covers larger area and high density of charged particles. Moreover, the most convenient feature
of microwave-assisted high-pressure NTP is efficient power coupling from the generator to plasma,
reduced flow rate of gas, and it also prevent electrodes from contamination. This microwave discharge
plasma can be applied for sterilization, decontamination of gases, treatment of surface, lightning, and
elemental analysis etc. (Tsuji et al., 2008; Quan et al., 2015).
Pulsed discharge plasma: The efficiency of non-thermal pulsed plasma depends on factors like
fast-rising voltage, short pulse width and long life capacity of pulsed power source. It has recently
become more popular due to its simplicity. The application of pulsed discharge plasma has high
removal efficiency, low cost, and economic use in various military, industries, and environmental
operations. It is also applicable for the treatment of water and gas, sterilization of medical appli-
ances, growth of plants and seed germination, etc. (Smulders et al., 1998; Wanga et al., 2010).
15.4 pLaSMa IN FOOD prOCESSING
15.4.1 Microbial Inactivation
Considerable research work has been done in recent past to determine the decontamination
mechanism of plasma (Table 15.1). The sterilization effect of plasma was first documented in 1960
and the patent was granted for plasma-based sterilization method in 1968 (Menashi, 1968). On the
table 15.1 application of Non-thermal plasma to reduce Microbial Load in Food Items
reduction
of Microbial
treated plasma Source/ Count
S. No. Sample Microorganism tested process Gas (log unit) references
1 Apple E. coli o157:H7, Gliding arc dried, 2.9–3.7 niemira and
S. stanley filtered air sites (2008)
2 Apple, E. coli o157:H7, DbD 1–35 Critzer et al.
Cantaloupe Salmonella spp., (2007)
melon skin L. monocytogenes
3 Cherry tomato E. coli o157:H7 DbD, air up to 4 schwabedissen
strawberry, et al. (2007)
iceberg
lettuce
4 spinach, S. typhimurium Lt2 nonthermal 3–4 Zhang et al.
lettuce, low-pressure (2013)
tomato, and oxygen plasma
potato (March Cs-1701
reactive ion
etching system)
5 Melon skin, E. coli, G. liquefaciens, Plasma jet, He+o2 1–3 Perni et al.
Mango skin P. agglomerans, (2008a)
S. cerevisiae
6 Melon flesh, E. coli, G. liquefaciens, Plasma jet, He+o2 1–2.5 Perni et al.
Mango flesh L. monocytogenes, (2008b)
S. cerevisiae
7 spinach E. coli O157:H7 DbD, air, o2 up to 5.8 Klockow and
Keener (2009)
8 radicchio (red E. coli, L. DbD 1.35–2 Pasquali et al.
chicory) monocytogenes (2016)
9 Fresh corn E. coli APPJ/Argon 2.1–3.6 baier et al.
salad leaves (2013)
10 orange juice Staphylococcus aureus, DbD, air 5 shi et al. (2011)
E. coli, Candida
albicans
11 Apple juice E. coli o157:H7 needle/plate up to 7 Montenegro
system, submersed et al. (2002)
12 Apple Juice Citrobacter freundii APCPJ/Argon, up to 5 surowsky et al.
Argon+o2 (2014)
13 Liquid media E. coli AtCC 25922 DbD-ACP/Air up to 7 Ziuzina et al.
(phosphate (2012)
buffer
solution)
14 sweet pepper Pantoea agglomerans DbD, He+o2 0.8–2 Vleugels et al.
(2005)
15 Almonds E. coli DbD, air 1.8–5 Deng et al.
(2007)
16 Almonds Salmonella, E. coli Dry air, Dry n2 1.14–1.34 niemira (2012)
o157:H7
17 Peanuts, A. parasiticus Low-pressure 1.8–5 basaran et al.
Hazelnuts, plasma, air, sF6 (2008)
Pistachio nuts
18 Cereals grains A. parasiticus, Low-pressure 1.8–5 selcuk et al.
Penicillium Ms1982 plasma, air, sF6 (2008)
19 brown rice A. flavus Cold APPJ/Argon no suhem et al.
cereal bars germination (2013)
(Continued)
table 15.1 (Continued) application of Non-thermal plasma to reduce Microbial Load in Food Items
reduction
of Microbial
treated plasma Source/ Count
S. No. Sample Microorganism tested process Gas (log unit) references
20 Processed L. monocytogenes needle/plate 0.2–1.7 song et al.
ham slices system, He (2009)
21 Chicken L. monocytogenes Plasma jet, He, n, 1.3–6.5 Lee et al.
breast, Ham o2 (2011)
22 Chicken breast S. typhimurium APPJ/n2+o2 0.66–1.33 Kim et al.
and pork loin (2013b)
23 raw chicken E. coli APPJ/n2+o2 1.53–1.76 yong et al.
breasts (2014)
24 Pork butt and L. monocytogenes, DbD plasma/ 1.90–2.68 Jayasena et al.
beef loin E. coli o157:H7 and Atmospheric gas (2015)
S. typhimurium
25 Chicken breast Campylobacter jejuni DbD, Air 0.5–3 Dirks et al.
(2012)
26 bacon E. coli, L. needle/plate 1–4.6 Kim et al.
monocytogenes, system, He, He+o2 (2011)
S. typhimurim
27 egg (with S. enteritidis, DbD, air+H2o up to 4.5 ragni et al.
shell) S. typhimurium (2010)
28 Cooked egg L. monocytogenes APPJ/n2, He+o2, no viable Lee et al.
white and and n2+o2 count (2012)
yolk
29 Chicken meat, Listeria innocua DbD, o2, Ar up to 3.5 noriega et al.
Chicken skin (2011)
30 Cheddar E. coli o157:H7, DbD plasma/ 2.1–5.8 yong et al.
cheese slices L. monocytogenes Atmospheric gas (2015a)
and S. typhimurium
31 Pet foil B. subtilis Microwave power 106–107 schneider et al.
plasma/Air (2005)
other hand, the oxygen-based plasma was applied very first in 1989. A wide range of microorganism
including spores and viruses can be effectively inactivated using plasma treatment (Lee et al., 2006;
Terrier et al., 2009). Hury et al. (1988) reported that the plasma generated at 200 W is sufficient to
reduce the population of Bacillus subtilis up to 3.5 log cycles after 5 min of treatment. The lethal
effect of plasma on microbial cells is due to interaction between the cell and the plasma ions.
High-intensity radical bombardment by plasma on the surface of exposed microorganisms causes
lesions due to accumulation of electrostatic forces on the exterior of cell surface. These provoked
lesions are unable to be repaired quickly by the microorganisms leading to rapid destruction of
living cell. This phenomenon of destruction is called “etching”, which basically depends on inter-
action between the substrate molecules and highly reactive species and ions of plasma (Pelletier,
1992). The antimicrobial effect of plasma is mainly associated with the action of its reactive species
and charged particles. Reactive species such as ozone, singlet oxygen, super peroxide, hydrogen per-
oxide, and nitrous oxide along with UV radiation act on microbial cells (Gaunt et al., 2006). These
highly reactive species break chemical bonds and damage DNA while interacting with the cell
membrane. The interaction of reactive species with water molecules forms hydroxyl radicals which
are highly reactive, leading to harmful effect on cells (Zou et al., 2004). Dobrynin et al. (2009) also
reported potential mechanism behind microbial inactivation that includes the breakdown of chro-
mosomal DNA by the highly reactive species of applied plasma; in addition, this cell also undergoes
certain oxidation and peroxidation reactions. Microbial inactivation efficiency of plasma is moisture
dependent, which was observed higher during the treatment of moist biological cells compared
to lowest in dry cells. The formation of hydroxyl radicals is important as they are responsible for
damaging 90% of DNA by forming hydration layer around it. The hydroxyl radical formed during
plasma treatment also interacts with certain organic compounds of the cell leading to a chain of
oxidation-reduction reactions. These reactions cause breakdown of cellular membrane, DNA, and
other components, ultimately causing cell death. It is well documented that microorganisms are
highly susceptible for singlet oxygen species which are more highly reactive than molecular oxygen
(El-Aziz et al., 2014). The interaction of lipids, proteins, and DNA with singlet oxygen leads to
destruction of microbial cell. Singlet oxygen breaks the double bonds of lipid bi-layer and causing
imbalance in transportation of molecules inside and outside of the cell (Montie et al., 2002; Mogul
et al., 2003; Guzel-Seydim et al., 2004). As the plasma species interact with the substrate in different
ways (activating different pathways in different organisms), it is important to use it selectively for
inactivation of microorganism without affecting the quality of host.
15.4.2 Enzyme activity
Fruits and vegetables are more prone to secondary losses mostly due to enzymatic browning
after harvest and during storage. Peroxidase and polyphenol oxidase are the well-known endog-
enous enzymes in fruits and vegetables responsible for enzymatic browning and also leading to
off flavours by oxidizing the phenols at the expense of H2O2 (Surowsky et al., 2013). Heating,
blanching, sterilization, and chemical preservatives are some most common traditional methods
practiced to prevent occurrence of enzymatic browning during processing (Anthon and Barrett,
2002; Elez-Martinez et al., 2006). However, HP processing, PEF, and irradiation are some recently
applied unconventional non-thermal techniques for inactivation of these endogenous enzymes
(Thakur and Nelson, 1998; Giner et al., 2001; Zhang et al., 2006; Zhong et al., 2007).
Recently, it was observed that application of plasma is not only effective for microbial inactiva-
tion but it also facilitates inhibition of simple biological compounds like enzymes in food products
(Table 15.2). The considerable reduction was observed in enzymatic activity of trypsin (zero at
4 J cm−2) due to application of plasma. The mechanism behind plasma inactivation of enzymatic
activity is breakdown of peptide bonds and denaturation of 3D structure of proteins in trypsin.
The investigation on effect of CP treatment of guava pulp and the whole fruit shows 70% and 10%
reduction, respectively, in polyphenol oxidase activity (Dobrynin et al., 2009). The kinetic study
shows decrease in peroxidase activity in tomato by the application of atmospheric air DBD plasma
of different voltage (Pankaj et al., 2013). Changes in metabolic activity of fresh cut apples were
determined after atmospheric pressure DBD plasma treatment over a time interval of 10, 20, and
30 min. The result shows highest reduction in browning area, i.e., 65% after 30 min of treatment, as
compared to control samples. Moreover, polyphenol oxidase residue was decreased eventually up
to 42% with increasing treatment time. The result shows that reduction in browning area in plasma-
treated apple slices is due to reduced metabolic activity as compared to control samples (Tappi
et al., 2014). In another study, activity of polyphenol oxidase and peroxidase enzymes in apples and
potatoes were also reduced remarkably by the treatment of plasma-processed air. The treatment of
apple and potato slices shows 62% and 77% reduction in polyphenol oxidase activity, respectively,
whereas activity of peroxidase enzyme was reduced up to 65% in cut apple and 89% in potatoes
(Bubler et al., 2017).
15.4.3 Starch Modification
Treatment of starch granules through plasma changes their physical properties such as alteration
in their structure, morphology, and rheology (Bie et al., 2015). The treatment of plasma leads to dry
etching on the surface of starch granules and ultimately modifying the properties (Pashkuleva et al.,
2005). The hydrophilicity of surface of starch granules increases with increasing interaction between
table 15.2 application of Non-thermal plasma for Functional Modification of Food properties
plasma Source/
S. No. Food Items process Gas Functional Modification references
1 enzyme CP/Air reduced polyphenol oxidase and surowsky et al.
peroxidase enzyme activity (2013)
2 tomato DbD atmospheric Decreased peroxidase enzyme Pankaj et al. (2013)
pressure CP activity
3 Apple slices Dielectric barrier Decreased polyphenol oxidase tappi et al. (2014)
discharge/Air activity
4 Fresh-cut kiwi fruit Microwave- Prevented colour changes and ramazzina et al.
assisted plasma darkening during storage (2015)
air/Humid air
5 blueberry DbD/Air reduced pesticide from the surface sarangapani et al.
(bocalid-80.18% and (2017)
imidacloprid-75.62%)
6 Pomegranate juice Atmospheric increased the yield of Anthocyanin bursac et al. (2016)
pressure CP (21%–35%)
reduction in colour
7 naringin Atmospheric increased radical scavenging Kim et al. (2014b)
pressure DbD activity, total phenolic content,
plasma and tyrosinase inhibition capacity
8 sour cherry Cold APPJ/Argon Higher retention of both Garofulic et al.
Marasca (Prunus anthocyanin and phenolic acids, (2015)
cerasus var. increased phenolic content
Marasca) juice
9 Wheat seeds surface discharge increased rate of germination Dobrin et al. (2015)
plasma/Air
10 Wheat flour Atmospheric enhanced dough strength and Misra et al. (2015)
pressure CP/ optimum mixing time, improved
Humid air viscous and elastic moduli
11 Wheat protein CP/oxygen Modified the structure of Protein, segat et al. (2015)
isolate reduced foaming, and emulsifying
capacity
the surface and oxygen-containing groups (OH, COOH, and CHO). Depolymerization and cross
linking of starch molecules could be a mechanism responsible for their modification in which two
starch molecules get rearranged by cross linking and releases water molecule by the breakdown of
glycosidic bonds are present near the amylopectin side chain (Table 15.2). This has resulted in water
molecule dissociates into its atoms and leaving no residual compound (Perez and Lares, 2006).
Plasma treatment enlarges the channels, reducing the degree of crystallinity, and forms car-
boxylic groups by partially oxidizing hydroxyl groups in starch molecules. These changes are
due to characteristic structure of starch granule having pinholes, allowing plasma to enter their
interior. Degradation of starch molecules by the plasma treatment causes changes in rheologi-
cal properties of starch paste by influencing their flow properties resulting in reduced viscosity
and increased starch concentration. The alteration in functional properties of starch granules
improves tensile strength and mechanical properties in starch films. Modification of starch using
natural polymers is sensitive towards etching particularly when it comes in contact with strong
acids. Therefore, conventionally week chemicals are used to alter the surface properties of starch
(Bie et al., 2015).
15.4.4 Seed Germination
Laboratory-scale observation shows that early germination of seeds is possible by plasma

treatment. The active particles of plasma directly penetrate through the seed coats causing
changes in cell structure. The possible mechanism behind increase in germination rate is due
to etching phenomenon of plasma. Permeability of seed coat increases by the etching action of
plasma, leading to enhanced transmission rate of oxygen and moisture to the embryo. Fridman
(2008) reported that plasma treatment increased the rate of seed germination by breaking their
dormant stage. The oxidative nature of plasma species damage cellular components including
DNA and cell wall, which may induce natural signals to increase the production of growth fac-
tors, modification of protein structure, and rate of enzymatic activity causing breakdown of seed
dormant stage.
The rapid germination and early seedlings in plasma-treated brown rice are likely to be triggered due
to increased enzymatic activity. Chen et al. (2016) observed that plasma treatment increased the level of
gamma-amino-butyric acid from 19 to 28 mg/100 g. The rate of enzyme mainly α-amylase activity was
also increased significantly in treated groups than in control during germination of brown rice.
Dobrin et al. (2015) reported that the surface barrier discharge treatment marginally affected
the rate of wheat seeds germination, whereas the growth parameters were influenced profoundly.
Investigation shows that roots and sprouts of germinated wheat seed were longer and heavier than
control samples when treated with plasma. The moisture absorption capacity of treated samples was
increased which might be a potential reason of improvement. Dhayal et al. (2006) also mentioned
that the reactive species of plasma changed surface properties of seeds and resulted up to 50%
increase in the rate of wheat seed germination.
15.4.5 Functional properties of Food
It is of great interest to distinguish and understand the effect of plasma species on bioactive
properties of foods during processing in order to avoid nutritional losses and determine the rele-
vance of plasma treatment for future applications. The antioxidant activity of bio-active compounds
protect cells from damaging action of free radicals like hydroxyl ions, singlet oxygen, superoxide,
peroxy radicals, peroxy nitrite, etc. (Percival, 1996).
There is great complexity in understanding the effect of plasma processing on sensory and
changes in bioactive component. Grabowski et al. (2016) studied characteristic changes, i.e., sensory
and phenolic content in spices by the treatment of low-pressure CP. The result showed considerable
reduction in intensity of aroma of almost all spices as well as change in colour and polyphenols
content of cayenne and sweet pepper. The treatment increased polyphenols content in case of cin-
namon, black pepper, marjoram, basil, and rosemary, whereas, in some spices such as thyme, sweet
and cayenne pepper, and ginger, the polyphenol content decreased. The study suggests that the
treatment of spices with CP under reduced pressure has negative effect as it reduces the charac-
teristic aroma of spices. Kim et al. (2014a) reported increase in radical-scavenging activity, FRAP
value, and total phenolic content of naringin (i.e., from 1.45% to 38.20%, from 27.78 to 207.78 μM/g,
and from 172.50 to 225.83 ppm, respectively) when given plasma treatment for 20 min. Garpfulic
et al. (2015) also reported increase in anthocyanin and phenolic content of sour cherry Marasca
juice by the treatment of atmospheric pressure plasma. They suggested that this increase in amount
of phenolic content could be due to dissociation of undefined small-sized aggregates or particles by
the plasma treatment.
15.5 appLICatION OF pLaSMa IN FOOD prOCESSING INDUStrIES
The aim of plasma treatment is to enhance the food safety by reducing microbial count as
much as possible without affecting physico-chemical quality and sensory attributes of food.
In recent years, plasma treatment has been investigated successfully for microbial decontami-
nation in the case of food and packaging materials, e.g., Escherichia coli in fresh produce
(Bermudez-Aguireet et al., 2013); Erwinia carotovora in potatoes (Moreau et al., 2007); Aspergillus
and Penicillium species in cereals, legumes, and vegetables (Selcuk et al., 2008); Listeria monocy-
togenes from aluminium foil, paper cups, and plastic trays (Yun et al., 2010), etc. NTP also facili-
tates several advantages over TP and other conventional technologies such as low temperature, less
cost, reduced heating losses, and high removal efficiency. Properties like low temperature and high
microbial inactivation efficiency are the most attractive features. NTP facilitates a wide range of
opportunities including microbial inactivation and surface decontamination of almost all food items
like meat, poultry, dairy, freshly harvested horticulture produce, and sprouted seeds. It can be also
employed successfully for sterilization and modification of packaging material imparting desired
functional properties in food packaging (Misra et al., 2015; Pankaj et al., 2015; Scholtz et al., 2015).
There are various laboratory and industrial scale studies conducted by researchers showing plasma
as a potential non-thermal technique useful in processing of various food items including cereals,
pulses, fruits and vegetables, spices, milk, meat, poultry, etc.
15.5.1 application in Cereal Industry
Cereals are widely grown agriculture crops, covering more than 50% of human consumption
(Poutanen et al., 2014). Recently, plasma is being used as a most promising cold processing technol-
ogy in cereals industry for the treatment of various crops. In addition to decontamination, plasma
processing also offers various valuable benefits for cereal industry including physico-chemical
modification of seed, water absorption capacity of seed, increased seed germination rate, preven-
tion from attack of insects during storage, etc.
In food industries, starch is utilized in many food preparations as a food additive. The altera-
tion in functional properties of starch granules improves tensile strength and mechanical prop-
erties in starch films. Starch is sensitive towards etching; when it comes in contact with natural
polymers particularly with strong acids, their functional properties get affected. Therefore, con-
ventionally week chemicals are used to alter the surface properties of starch. Pashkuleva et al.
(2008) reported that dry etching property of CP is effective for modifying the surface and surface
cleaning of bio-polymer. The hydrophilicity of starch granules surface increases when introduc-
ing oxygen-containing groups like OH, COOH, and CHO groups. Depolymerization and cross
linking of starch molecules could be a mechanism responsible for this type of modification. Perez
and Lares (2006) reported that in depolymerization two starch molecules get rearranged by cross
linking due to breakdown of glycosidic bonds present near the amylopectin side chain and releas-
ing water molecule. This resulted water molecule dissociates to its atoms, leaving no residual
compound. Misra et al. (2015) studied the treatment of atmospheric plasma to investigate changes
in rheological properties of both soft and hard wheat flour. The investigation shows improvement
in rheological properties such as dough strength and optimum mixing time of both soft and hard
wheat flour. The rate of changes in rheological properties varies with the range of treatment time
and power applied. Increase in time and power of applied plasma treatment progressively changes
the elasticity and viscous moduli of strong wheat flour. Fourier transform infrared spectroscopic
(FTIR) evaluation showed increase in α-helix and β-turns and decrease in β-sheets in second-
ary structure of proteins for both week and hard wheat flour, leading to alter their functional
properties.
Mir et al. (2015) reported that brown rice is a rich source of nutrients; however, its reduced desir-
ability is due to poor cooking and eating quality. Chen et al. (2012) reported that plasma discharge
is useful in modifying the properties of brown rice. They used low-pressure plasma to improve
the quality characteristics such as cooking, microstructure, and textural properties of brown rice. The
phenomenon of etching changed the surface properties of treated brown rice and improved water
absorption capacity of soaked rice kernels. The treatment also reduced cooking time and softens
the texture resulting improved chewing properties of brown rice. The result shows that increase
in number of iodine binding sites in structure of brown rice kernel is due to plasma treatment.
Lii et al. (2002) reported that plasma treatment increased the availability of amylose for iodine
reaction thereby resulted in increase in shifting in wavelength in UV-VIS band. Perez and Lares
(2006) reported rearrangement of intermolecular bonds lowering the water absorption capacity due
to breakdown of starch molecule. Szymanowski et al. (2005) studied the effect of RF plasma on
properties of treated potato starch films. The treatment of plasma results in de-agglomeration of
starch granules by reducing the formation of capillaries. Plasma-treated starch showed separate
granules, whereas agglomeration was observed in untreated samples.
Plasma processing can be also used for green production of starch products that bear low vis-
cosity at a relative high concentration. The modification of starch through plasma treatment is an
important tool for altering its physical structure and rheological properties. Bie et al. (2015) reported
significant effect of DBD plasma on morphology, structure, and rheological properties of treated
corn starch. Application of DBD plasma is not only limited to the surface of treated starch granules
but also it enlarges the channels, reduces the degree of crystallinity, forms carboxyl groups by par-
tial oxidation of hydroxyl groups, and degrades molecules. These changes are due to characteristic
structure of starch granule having pinholes, allowing plasma to enter their interior. Degradation of
starch molecules during processing is responsible for changes in rheological properties of starch
paste from non-Newtonian to Newtonian fluid, reduced viscosity, and increased starch concentration.
Sarangapani et al. (2015) investigated changes produced in cooking and textural charac-
teristics in parboiled rice treated with low-pressure plasma. The moisture absorption capacity
of treated parboiled rice increased; however, cooking time reduced up to 8 min. The textural
characteristics of rice like hardness and stickiness were also decreased with increasing treat-
ment power and time.
Plasma is also effective in reducing microbial count of cereals without or only marginally
affecting their inherent characteristics. Seeds of wheat, rye, oat, barley, corn, bean, chickpea, and
lentil were decontaminated successfully by reducing the strength of contaminating Aspergillus par-
asiticus and Penicillium sp. to less than 1% of their initial count (Selcuk et al., 2008). Application
of plasma on the surface of brown rice bar reduced growth of Aspergillus flavous up to 4 cfu/g of
bar. The reduction was achieved by exposing the surface of bar for 20 min to a power source of
40 W plasma. This treatment prevented growth of Mycelium on the surface of bars for at least 20 days
(Suhem et al., 2013). Plasma processing is also effective for inactivation of bacterial endospores in
treated cereals. Exposure of contaminated wheat grain to atmospheric DBD plasma resulted inac-
tivation of bacterial endospores. This inactivation of endospores is due to defects produced by the
impact of highly reactive plasma species containing ions and singlet oxygen. The efficiency of
plasma treatment varies with treatment variables like treatment time, pulse frequency, and voltage
applied during processing (Butscher et al., 2016).
Storage of seasonal horticulture produce including cereal crops is one of the major steps in
postharvest management for their sustained utilization and processing throughout the year. There
are several obstructions like attack of insects, rodents, termites, moths, etc. affecting the shelf
life and quality of cereals during storage. The intensity of plasma jet pulses and distance between
the object and the nozzle of plasma jet are two processing variables used for the treatment of
moth Plodia interpunctella (El-Aziz et al., 2014). The research shows a significant increase in
mortality of larvae and pupae as well as decrease in emergence of adults. The observation sug-
gests that mortality of larvae and pupae and reduction in emergence of adults is proportional to
intensity of applied plasma pulses but indirectly proportional to distance from the nozzle. This
inactivation of moths is possibly due to generation of high oxidative stress in their body caused
by the reactive species of NTP. These studies suggest that application of atmospheric plasma is
useful in processing, preservation, and modification of cereals and related products. In addition,
it is comparatively less expensive and/or potential alternative of other non-thermal methods for
enhancing the quality.
15.5.2 Seed Germination
Laboratory-scale investigations show early germination of seeds is possible by treatment of

plasma. The active particles of plasma directly penetrate through the seed coats and resulting
changes in cell structure. The possible mechanism behind increase in germination rate is supposed
to be etching phenomenon of plasma. Permeability of seed coat increases by the etching action
of plasma leading to enhanced transmission rate of oxygen and moisture to the embryo. Fridman
(2008) reported that plasma treatment increased the rate of seed germination by breaking their dor-
mant stage. The oxidative nature of plasma species damaged cellular components including DNA
and cell wall. They suggested that the treatment of plasma may induce natural signals to increase
the production of growth factors, modification of protein structure, and the rate of enzymatic activ-
ity which could also be associated with early breakdown of seed dormant stage.
Chen et al. (2016) reported increase in seedling length, germination percentage, and water-
absorbing capacity of brown rice when treated with low-pressure plasma (1–3 kV for 10 min).
The treatment of 3 kV for 10 min of plasma was found best among all the treatments. The rate of
enzyme mainly α-amylase activity was increased significantly in treated groups as compared to
control during germination of brown rice. The rapid germination and early seedlings in plasma-
treated brown rice are likely to be triggered due to increased enzymatic activity. They observed
that plasma treatment increased the level of gamma amino butyric acid from 19 to 28 mg/100 g.
Moreover, antioxidant activity was also increased significantly in treated brown rice compared
to controls.
Dobrin et al. (2015) studied the behaviour of wheat seeds treated with surface barrier discharge
plasma under ambient temperature and pressure. The treatment has marginal effect on rate of seeds
germination but the growth parameters were influenced profoundly. Investigation shows that roots
and sprouts of germinated wheat seeds were longer and heavier than control sample when treated
with plasma. The increase in moisture absorption capacity of treated samples might be a potential
reason of this improvement.
Jiafeng et al. (2014) reported 6.7% increase in germination rate of wheat grains when treated with
helium plasma. A significant increase in growth rate was observed in treated plants as compared to
control sample during field experiments. Dhayal et al. (2006) observed that rate of seed germina-
tion increased up to 50% when treated with CP. They also mentioned that reactive species of plasma
changed the surface properties of seeds which resulted in increase in rate of seed germination. Shao
et al. (2013) used arc plasma to determine its effect on germination and seedling growth of spin-
ach seeds. They observed increase in rate of germination by 137.2%, whereas germination strength
increased 217.6% by the treatment.
The rate of seed germination can be also delayed by the application of plasma treatment and can
be used for storage of seed over years. Volin et al. (2000) investigated delay in germination by the
treatment of CP in CF4 and octadecafluorodecalin-coated seeds including soy beans, pea, corn, etc.
They observed that the thickness of coating is directly associated with delay in seed germination.
The coating of octadecafluorodecalin is more effective for delaying the germination of seed than
samples coated with CF4.
Filatova et al. (2011) observed 10%–20% increase in germination of legumes during both
laboratory- and field-level investigations. Moreover, active species of plasma-like singlet oxygen
and hydroxyl radicals also function as a probable sanitizing agent that reduced fungal growth by
3%–15% in treated legume seeds.
15.5.3 herb/Spice processing Industry
Herbs and spices are used worldwide as an essential part of food formulation. They are well
known for their specific flavour, aroma, and colour and are rich source of bioactive compounds.
These are widely used for making variety of dishes in house hold as well as in commercial food
processing to give specific colour, flavour, and aroma. Most of the herbs and spices are traded
worldwide in their natural, dry, or powder forms. Their contamination basically depends on
hygienic practices at the time of harvesting and the drying process. Some herbs and spices may
contain microorganisms due to unhygienic practices during handling that can be pathogenic for
humans like Salmonella and Bacillus cereus (Mckee 1995). Buckenhuskes and Rendlen (2004)
reported increased food-borne diseases and intoxication over the last decade of twentieth century,
which was also related with the poor hygienic practices during handling of herbs and spices at
the time of harvesting. FDA (2013) reported that in 2010 alone up to 6.6% cases were observed,
where spices imported into United States and in other countries were infected with bacteria mainly
genus Salmonella. Gieraltowski et al. (2013) studied the causes of nationwide outbreaks. The study
shows that ready-to-eat (RTE) products are mainly involved in these outbreaks. Consumption of
RTE salami products which were added with contaminated black and red pepper after the con-
trol measures of pathogen reduction, resulted in nationwide higher outbreak of S. Montevideo
infection. Therefore, it is important to decontaminate raw spice to reduce the chance of contamina-
tion after safety measures.
There are a number of technologies currently in use for the decontamination of herbs and
spices such as application of ethylene oxide, gamma irradiation, or use of steam; but, they also
have some associated drawbacks. The decontamination efficiency of gamma irradiation is quite
efficient but consumers have a negative perception of irradiated products. In the European
Union, the use of ethylene oxide for fumigation of spices is prohibited due to its carcinogenic
potential in humans, although thermal inactivation process by using steam treatment is exten-
sively used in Europe. However, most of the herbs and spices contain compounds that are heat
sensitive leading to alteration of their inherent flavour and aroma (Schweiggert et al., 2007).
Suresh et al. (2007) conducted study on heat processing of various spices such as turmeric,
red pepper, and black pepper and reported significant losses in concentration of their bioactive
compounds. Grabowski et al. (2014) studied the sterilization possibilities of black pepper using
low-pressure CP as an alternative method. The study shows that O2-assisted plasma gave bet-
ter results than nitrogen, air, argon, and H 2O2 gases, and higher reduction of microbial load in
black pepper was observed. The treatment resulted in up to 1 log cycle reduction in both aero-
bic spore-forming and non-spore forming bacteria, and spore-forming anaerobic bacteria count
was undetectable after 60 min of treatment. The effect of microwave-powered CP treatment on
microbial load including Aspergillus flavus and Bacillus cereus spores of red pepper powder
was also investigated by Kim et al. (2014a). The result shows that 20 min of treatment decreased
up to 1 log CFU/g of naturally occurring total aerobic bacteria, where the power and pressure
of applied plasma were 900 W and 667 Pa, respectively. The population count of A. flavus was
reduced up to 2.5 log spores/g using same level of treatment. Whereas, the count of B. cereus
spores was reduced by 3.4 log spores/g when CP treatment was integrated with heat treatment
at 90°C for 30 min.
Hertwig et al. (2015a) used NTP treatment to determine its impact on natural microbial load
and colour changes in herbs and spices (pepper seed, crushed oregano, and paprika powder). The
treatment reduced up to 3 log cycles of microbial load present naturally in paprika powder and pep-
per seeds, after 60 min of exposure. The treatment of oregano resulted in less reduction of micro-
bial load, i.e., 1.6 log cycles. However, in red paprika powder considerable loss in colour was also
observed when treated for 5 min or more, whereas in the case of pepper seeds and oregano minor
impact on colour was observed.
Hertwig et al. (2015b) also investigated decontamination efficiency of two different atmospheric
pressure plasma discharge methods, i.e., microwave-driven remote plasma and plasma jet. Naturally
contaminated black pepper samples were further inoculated with known pathogens (i.e., Bacillus
subtilis spores, Bacillus atrophaeus spores, and Salmonella enteric) and treated separately with two
different plasma discharges. Microwave-driven remote plasma treatment was found more effective
in reducing the pathogen load as compared to direct plasma jet treatment. The reduction in pathogen
count of B. subtilis spores, B. atrophaeus spores, and S. enteric was achieved up to 2.4, 2.8, and 4.1
log cycles, respectively, after 30 min of treatment using remote microwave plasma.
15.5.4 Fruits and Vegetables processing Industry
Plasma is a most promising technology to improve quality and shelf life of agriculture produce.
There are several studies conducted over last decade for determining the applicability of plasma in
fruits and vegetables processing industry. Plasma processing is capable of reducing microbial load
from the food surface, due to highly reactive species, which enables it as an alternative method for
sanitizing the surface as well as processing and preservation of fruits and vegetables.
Basaran et al. (2008) investigated fungicidal effect of low-pressure CP using air and sulphur
hexafluoride gases. The treatment was carried out for 20 min duration using various nuts including
peanuts, hazelnuts, and pistachio nuts, contaminated with pathogen A. parasiticus. Higher reduc-
tion in pathogen count was observed during early period of treatment. Samples treated with air
plasma reduced 1 log cfu/g in early 5 min, but an additional 1 log cfu/g was reduced in further 5 min
of treatment. Although, observation shows that the treatment of plasma using sulphur hexafluoride
gas had higher impact on microbial count and resulted in five-fold more reduction, i.e., 5 log cfu/g
for the same period of treatment as in air gas plasma. They also reported that the air plasma treat-
ment is more efficient than sulphur hexafluoride gas plasma for inactivation of fungal toxin. The
treatment using air gas plasma reduced up to 50% of total aflatoxins (i.e., aflatoxin B1, B2, G1, and
G2), whereas sulphur hexafluoride gas plasma resulted in only a 20% reduction of total aflatoxin
after 20 min of treatment.
Niemira (2012) used atmospheric pressure CP treatment for decontamination of dry almonds
infected with Salmonella and E. coli species. The samples were treated to a time interval of 0,
10, and 20 s using variables dry air or nitrogen gas, where distance of the sample from the emit-
ting source was 2, 4, or 6 cm. Significant reduction was observed in both pathogens Salmonella
and E. coli. The reduction in E coli count was found to be higher, i.e., 1.34 log cfu/mL when
treated for 20 s at 6 cm spacing. The inactivation efficiency of plasma varies with duration of
treatment, distance of the sample from the plasma emitting source, and the type of pathogen
involved in process. Study shows less antimicrobial efficacy of N2 gas-assisted plasma than dry
air plasma-treated almonds. The effect of CP on Citrobacter freundii present in apple juice was
investigated by Surowsky et al. (2014). The treatment for 8 min using argon and 0.1% oxygen
gas plasma along with subsequent storage of 24 h reduced load of C. freundii in apple juice up
to 5 log cycles. The report suggests that for successful reduction in microbial population it is not
necessary to expose microbial cell directly to the plasma source. Generation of compounds like
hydrogen peroxide and hydroxyl radicals in liquid during plasma treatment are mainly responsi-
ble for inactivation of microbes. The reduction in enzymatic activity and browning area was also
observed in fresh cut Pink Lady apples by the treatment of DBD plasma (Tappi et al., 2014). The
fresh cut apple slices were exposed to plasma for a period of 10, 20, and 30 min and the qual-
ity parameters were assessed immediately after the treatment and during 24 h of storage. The
result showed significant reduction in browning areas, i.e., 65% by the treatment of 30 min along
with subsequent storage of 4 h. The activity of enzyme polyphenoloxydase also reduced linearly
up to 42% with increase in treatment time; however, other quality parameters were remained
partially affected by the treatment. The metabolic activity in tissues was reduced in plasma-
treated apple slices as compared to control. The effect of atmospheric pressure CP treatment of
in-pack cherry tomatoes was investigated by Misra et al. (2014b). The quality parameters like
change in respiration rate, weight loss, change in pH, and firmness were evaluated during storage
of DBD plasma-treated in-pack cherry tomatoes. Significant changes were observed in quality
parameters during storage except colour values and respiration rate of tomatoes. However, they
suggested that the application of CP effectively decontaminated and retained the quality of
cherry tomatoes.
CP could be an important alternative of chlorine washing in food processing industry, mainly
for large-scale decontamination of green leafy vegetables. Pasquali et al. (2016) studied the changes
in physico-chemical properties and microbiological quality of radicchio leaves immediately after
CP treatment and during storage. The colour and external appearance of treated radicchio leaves
were changed significantly during storage. However, antioxidant activity of radicchio leaves was
not affected by the exposure of 15–30 min in CP. The population of inoculated E. coli was reduced
significantly by the treatment of 15 min of CP; however, to achieve significant reduction in L. mono-
cytogenes count requires 30 min of exposure. The study suggests that decontamination of leafy
vegetables could be achieved effectively by optimizing conditions for CP treatment. Perni et al.
(2008c) used AC power discharge plasma to determine its effect on pericarp of melon and mangoes
inoculated by E. coli, G. liquefacien, P. agglomerans, and S. cerevisaes species. Significant reduc-
tion in microbial count was observed in both the samples; however, the treatment time and the rate
of inactivation varied with the type of species. Observation shows that microbial count of highly
resistant species, e.g., G. liquefaciens and P. agglomerans, were reduced below the detection limit
in both the samples after only 2.5 s of treatment, whereas inactivation of E. coli species required
5 s to reach the same level of reduction. Plasma treatment effectively decontaminated pathogenic
and spoilage causing microbes from the surface of treated fruits. Sarangapani et al. (2017) used
atmospheric CP to investigate dissipation efficacy of agrochemicals on blueberries. The treatment
of high-voltage DBD plasma on in-pack blueberries contaminated with pesticides (boscalid and
imidacloprid) was investigated. Plasma treatment with a power of 80 kV for 5 min was reduced the
concentration of pesticide on the surface of in-pack blueberries up to 80.18% of boscalid and 75.62%
of imidacloprid. However, total phenolics and flavonoid contents of blueberry were increased sig-
nificantly after 1 min of treatment for all applied voltages. The treatment for longer time shows
significant reduction in ascorbic acid content of blueberries. The changes in physical parameters,
mainly colour of blueberry, were not affected by the treatment, whereas change in firmness was
under the limit of acceptance.
15.5.5 Milk processing Industry
Raw milk is highly nutritious dietary supplement for good health of humans but on the other
hand it is also suitable for microbial growth. Application of plasma has an important role in disin-
fecting the heat sensitive materials in milk without increasing operational temperature. Therefore,
plasma processing has ability to sanitize raw milk and milk products without changing their natural
composition. Korachi et al. (2015) studied the effect of CP technology on biochemical properties of
whole raw milk samples, where milk was treated to a time interval of 0, 3, 6, 9, 12, 15, and 20 min.
The result showed significant changes in organic composition; however, the composition of lipid in
raw milk remained unaffected by the treatment. Increase in treatment time significantly increased
the level of total aldehyde content of milk, whereas the levels of total ketone or alcohol were not
changed significantly. Gurol et al. (2012) investigated the efficacy of low-temperature plasma for
sanitizing milk samples of different fat concentrations. The samples, i.e., whole, semi-skimmed,
and skimmed milk, were dispersed with E. coli before treatment with atmospheric plasma for spe-
cific time intervals of 0, 3, 6, 9, 12, 15, and 20 min. The observation showed that the level of fat
content in milk did not affect the treatment efficiency. The population of dispersed E. coli cells
significantly decreased up to 54% after 3 min of treatment. However, exposure of whole milk for
20 min of plasma reduced bacterial count 3.63 log cfu/g from their initial count of 7.78 log cfu/g.
Microbial growth was not observed during storage of 6 weeks, in addition to which colour and pH
of raw milk samples also remained unchanged by the treatment.
The effect of exposure time and power input of atmospheric plasma on microbial load of cheese
sample was investigated by Song et al. (2009). They reported decrease in rate of microbial count
with increasing treatment time and power input. Atmospheric pressure plasma was applied in sliced
cheese samples inoculated with cocktail of three-strain of Listeria monocytogenes. The treatment
reduced up to 8 log cycle in 120 s at 150 W. Thermal death value (D value) was calculated by plot-
ting the survival curves shows up to 90% reduction of microbial load when exposed to atmospheric
pressure plasma for 71.43, 62.50, 19.65, and 17.27 s and the power input was 75, 100, 125, and 150 W,
respectively.
Yong et al. (2015b) also used in-pack cheese slices (packed in rectangular plastic contain-
ers) to evaluate the changes in microbial load during storage after the treatment of encapsu-
lated atmospheric DBD plasma. The study reports that microbial load in sliced cheese was
decreased with increase in treatment time. The population of pathogenic microbes, e.g., E. coli,
S. typhimurium and L. monocytogenes reduced 2.67, 3.10, and 1.65 decimal with the treatment
time 60 s, 45 s, and 7 min, respectively. They also observed further decrease in pathogen load
of sample during post-treatment storage period. The study suggested that plasma treatment of
canned cheese slices was useful in preventing pathogens growth during post-treatment storage
and to enhance the shelf life.
15.5.6 Meat and poultry processing Industry
Raw and processed meats as well as poultry products are highly associated with number of
food-borne pathogens. The most frequently observed food-borne illnesses are mainly with the
consumption of unhygienic meat or egg contaminated by Salmonella spp., Listeria monocyto-
genes, Campylabacter spp., Clostridium spp., E. coli, and other enterohemorrhagic E. coli spp.
(Loretz et al., 2010; Kim et al., 2011). Microbiological contamination of meat and meat products
can occur during any stages of processing chain including preparation, storage, and distribution,
leading to food-borne illness in consumers. The challenge arising in decontamination of meat and
meat products are due to composition of meat which makes it highly perishable. Application of
conventional thermal methods of treatment is efficient in inactivation of food-borne pathogens,
but they also associated with some nutrient losses, texture losses, and reduction of meat sensory
properties. Therefore, plasma processing is being introduced as a potential non-thermal treatment
to achieve the microbiologically safe and wholesome foods. It is advantageous for meat process-
ing industries that most of the plasma discharge systems used in processing work at atmospheric
pressure and are cost effective. Highly reactive species of plasma, e.g., free radicals, electrons,
ions, and UV photons are well recognized for their microbial inactivation properties (Kaushik
et al., 2013). Plasma treatment was studied for its microbial inactivation efficiency along with
effect on nutritional attributes of in-pack fresh pork and beef samples (Jayasena et al., 2015). The
samples (pork-butt and beef-loin) were inoculated with L. monocytogenes, S. Typhimurium, and
E. coli O157:H7 and packed separately in a flexible food package system which was modified to
generate DBD plasma within the food package. The samples were treated for a period of 0, 2.5, 5,
7.5, and 10 min using thin-layer DBD plasma. The microbial counts of samples were reduced sig-
nificantly; however, significant increase in redness of meat was observed after 5.0 and 7.5 min of
exposures. Oxidation of lipid was influenced significantly only after 10 min of treatment. Texture
and sensory quality of treated and untreated meat samples were comparable, although minor nega-
tive changes in taste were observed in treated samples. Kim et al. (2011) evaluated the effect of
plasma gas composition (e.g., He and He+O2 gas mixture) on microbial inactivation efficiency.
The higher reduction of microbial count (L. monocytogenes, E. coli, and S. typhimurium) was
achieved by the application of He+O2 gas (e.g., 2–3 log cycles) mixture as compared to only He gas
(e.g., 1–2 log cycles) assisted plasma. Physico-chemical properties of treated bacon sample were
not changed significantly except the total colour value (L* value), which increased significantly
after the treatment. Kim et al. (2013a) further studied changes in microbial load of Salmonella
typhimurium in raw chicken breast and pork loin samples using APPJ under varied conditions,
including distance, time, and direction of the treatment. The treatment of samples on both sides for
5 min at a distance of 20 mm significantly reduced the count of S. typhimurium in treated chicken
breast and pork loin sample, i.e., 0.66 log CFU/g and 1.33 log CFU/g respectively. Yong et al.
(2014) also reported higher reduction in E. coli count when sample (raw chicken breast) was treated
both side at a distance of 20 mm. They found that the inactivation efficiency of plasma varies with
change in initial concentration of microbial count.
A significant quality change along with reduction in pathogen count was observed in pork loin
when treated with He and He+O2 gas-assisted DBD plasma. The reduction in E. coli count was
observed by 0.26 and 0.50 log cycles when treated with He and He+O2 gas-assisted DBD plasma for
5 min respectively; however, 0.34- and 0.55-log cycle reduction was observed after 10 min of treat-
ment, respectively. The treatment also shows 0.17 to 0.35 and 0.43 to 0.59 log cycles reduction in
L. monocytogenes when He and He+O2 assisted DBD plasma was applied to the samples for 5 min
and 10 min, respectively. The treatment also influenced some physico-chemical parameters like pH,
total colour value, and sensory quality (colour, odour, appearance, and acceptability, etc.) in pork
loin sample. Lipid oxidation was greatly affected by the He+O2 gas-assisted DBD plasma-treated
samples than other samples (Kim et al., 2013b).
Washing and cleaning of egg shells before or after grading is banned in the European Union.
Therefore, the need of alternative methods is raised in poultry processing industries. Ragni et al.
(2010) studied the application of non-thermal gas plasma for sanitizing the surface of eggs shell.
The samples containing pathogens S. enteritidis were exposed to gas plasma for a period of 0, 10,
20, 30, 45, 60, and 90 min to determine the sanitation efficiency. The result showed that the sanita-
tion efficiency of plasma varies with changing treatment time and relative humidity (RH) of the
samples. Population count of S. enteritidis was reduced up to 1.6 log CFU/egg shell after 10–20
min of treatment. A maximum reduction of 2.2–2.5 log/egg shell was achieved after 60–90 min
of exposure at RH 35%, although, treating the samples for 60–90 min at RH 65% resulted higher
reduction in population count, i.e., 3.8–4.5 log/egg shell. The effect of APPJ treatment on micro-
biological and physico-chemical quality of cooked egg white and yolk was studied by Lee et al.
(2012). Cooked egg samples were treated with APPJ using different gasses, such as N2, He+O2,
and N2+O2, to reduce microbial load. The population count of L. monocytogenes reduced up to
5 log cycle by the treatment of He gas plasma; however, pathogen count was undetectable when
treated with He+O2 and N2+O2 gas assisted APP for 2 min. The viable count of treated cooked egg
white sample increased approximately 3 log cycles after one week of storage, whereas there was
no significant viable count observed in egg yolk. The instrument colour value (e.g., L* value) was
decreased significantly by the treatment of both cooked egg white and yolk on day zero. Moreover,
some significant changes in sensory characteristics like flavour, taste, and overall acceptability were
also observed in APP-treated cooked egg yolks; however, treatment did not significantly affect the
sensory characteristics of cooked egg white.
15.5.7 application in Food packaging Industry
Packaging plays vital role in food industries; packaging materials are used for protecting food
from outside environment during handling, transportation, and distribution. Number of packag-
ing materials like metals, glass, paper, and polymers are used for packaging of various goods
depending on their suitability and functionality. Over the past few decades, polymeric packag-
ing materials have gained growing interest in various food processing industries due to their
physical and chemical nature. Polymeric materials serve various functions such as low cost, high
flexibility, less weight, required transparency, hydrophobic nature and can provide packages of
adequate strength (Medard et al., 2002; Vesel and Mozetic, 2012). In addition to this, properties
of polymeric materials can be improved by means of physical or chemical treatments resulting in

surface deposition, surface cleaning, or etching phenomenon. The purpose of treatment is to get
packages of desired properties like required water vapour transmission rate (WVTR), gas barrier
properties, removal of unwanted materials and contaminants, etc. Conventionally, there are vari-
ous physical and chemical techniques are employed in packaging industries for the surface modi-
fication of packaging material that includes UV light, irradiation, flame and corona treatment,
low-pressure plasma, etc. However, due to greater precision, an easy-to-control the process and
environmentally friendly nature, physical techniques are preferred to their chemical counterparts
(Adler et al., 1999).
CP technique has been recently applied in food packaging industries and gaining preferences
for improving barrier properties as well as removal of contaminants from the surface of pack-
aging material. It involves various physico-chemical interactions with the packaging polymers to
improve their functional properties. The barrier properties of packaging materials against gases
(O2, CO2) and chemical solvent get improved by the plasma deposition of barrier layer on the pack-
aging surface (Table 15.3). It also improves resistance to mechanical damage as well as sealing and
printing ability of packaging polymers (Schneider et al., 2009; Pankaj et al., 2014). In addition to
this, Muranyi et al. (2007) reported that plasma treatment can be used for sanitizing the packaging
surface within a short treatment time.
Almost all food safety and regulatory guidelines insist on the requirement of microbiological
safety of food packaging material. In case of HACCP system, packaging processes are consid-
ered as a critical control point. Sterilization of packaging materials has an important aspect in
food industries; if sterilization is not proper it could contaminate in-pack food items and can cause
greater health risks and economic losses. Many physical and chemical methods such as heating,
UV treatment, chemical treatments like ethylene and H2O2, etc. are used conventionally for ster-
ilization. Increase in overall processing cost and formation of liquid effluents by the conventional
methods of package treatment are major constraints for food industries. However, the use of plasma
is a well-recognized technique for effective sterilization of packaging material without leaving any
residues after treatment. It is a chemical-free, fast, and safe approach and can be applied to a wide
range of packaging materials (Schneider et al. 2005). Packaging materials such as packaging film,
plastic bottles, and lids can be sterilized effectively by the plasma treatment without changing their
bulk properties. Plasma assures negligible chances of shadowing effect as it flows all around the
packaging surface. The treatment of plasma assures low-temperature sterilization of heat sensitive
packaging materials such as polyethylene and polycarbon packaging without affecting material
properties (Shakila et al., 2012).
Keener et al. (2012) utilized DBD plasma for the treatment of various foods within the food
package using high-voltage electrodes. The gases contained within the package get ionized due to
high-voltage process during applied electric field. Temperature of the product surface increases to a
slighter extent when the reactive molecules are generated in significant amount during ionization. The
air present inside a 4 L re-sealable plastic (LDPE) bag required only 40–50 W of power for ionization.
Type of product, packaging material, composition of gas and configuration of package/electrode are
commonly affecting the specific treatment time required for the destruction of targeted spores and
bacteria. Recent advances in the system facilitate less processing time for the treatment of packages
filled with air having electrode gap of up to 10 cm (Keener et al., 2012). LDPE, HDPE, PETE, glass,
cardboard, rubber, polystyrene, and others are the most frequently used packaging materials for in-
pack ionization process.
Oh et al. (2016) studied the possibilities of CP treatment using various gases (O2, N2, air, He,
and Ar gas-assisted CP) to improve the quality of defatted soybean meal (DSM)-based edible film
for food. The results showed that CP-treated DSM films were easily decomposable. However, their
moisture barrier properties, tensile strength, and elongation properties were increased up to 24.4%,
6.8%, and 13.4%, respectively, by the treatment under optimal conditions, i.e., 15 min at 400 W.
table 15.3 application of Non-thermal plasma on Modification of packaging polymers

packaging plasma Source/
S. No. polymer process Gas Functional Modification references
1 Potato starch film Air Plasma increase in tensile strength, starzyk et al. (2001)
decrease in hydrophilic nature
2 boPP film rF Air increase in roughness ageing Mirabedini et al.
plasma/10–50 W affect, decrease in contact (2007)
angle
3 PP film Air Corona/30 kHz, increase in adhesion, decrease Dixon and Meenan
1.7 J/cm2 in contact angle (2012)
4 Pet film Glow discharge/10 increase in roughness, Pandiyaraj et al.
W crystallinity, and yield (2008)
degradation, decrease in
contact angle
5 PP film Diode plasma increase in surface energy, slepicka et al. (2010)
discharge/8.3 W decrease in contact angle
6 Pet film Jet plasma DC increase in hydrophobic nature Akishev et al. (2008)
discharge/35 W
7 Pet film Jet plasma/285 W increase in weight, decrease in Gotoh et al. (2011)
contact angle and wettability
8 HDPe film rF Aro2 increase in roughness, decrease banik et al. (2002)
Plasma/150 W in crytallinity, contact angle
9 LDPe film rF Aro2 increase in crystallinity, Ataeefard et al.
Plasma/25–100 W decrease in contact angle and (2009)
ageing effect
10 Pet, PtFe, DC Ar plasma/10 W Decreased contact angle, weight slepicka et al. (2013)
HDPe and PLLA of polymers and improved
film polymer cytocompatibility and
biocompatibility
11 Porous rF Co2 Plasma/40 increased hydrophilic nature, Patra et al. (2015)
polycaprolactone W increased pore size and crystal
scaffold form transition
12 Pe films APPJ He or He+o2 Changed in surface wettability, Jie and yiping (2015)
Plasma/ 40 W improved surface chemical
reorientation
13 Pet film rF o2 Plasma/200 increased hydrophilicity, Junkar et al. (2015)
W improved morphological
properties
14 Pet film rF H2s Large changes in surface Vesel et al. (2016)
Plasma/13.57 MHz morphology
15 Metal surface (Al, APPJ n2 and o2 Affecting surface cleaning and Kim et al. (2003)
sus and Cu Plasma/16–20 kHz particle aggregation, enhanced
metal) adhesion of surface, new
particles can be added to the
surface
Treated DSM film was also evaluated for packaging of smoked salmon, where it effectively reduced
hardness and lipid oxidation of smoked salmon during storage at 4°C.
15.6 FaCtOrS aFFECtING thE pLaSMa EFFICIENCY IN FOOD prOCESSING
Generation of plasma using low-pressure glow discharge is of great interest in fundamental

research. However, generation of low-pressure plasma requires costly air-tight enclosure which is
not feasible for industrial operation as it is highly expensive and time consuming. Therefore, vari-
ety of plasma sources developed that can operate under atmospheric pressure but retain all the
properties of low-pressure plasma (Nehra et al., 2008). There are several factors such as feed gas,
applied voltage, type of plasma discharge, type of food, and interaction plasma species with food
material, etc. affecting the efficiency of plasma (Mir et al., 2016).
15.6.1 Gas/Mixture of Gases
The efficiency of plasma directly associated with the type of operating gas used for discharge
generation such as air or nitrogen or a mixture of gases including neon, argon, and helium. The
mixture of gases may also include He/O2, He/O2/H2O, He/N2, Ar/O2, N2/O2, N2/N2O, etc. (Guo et al.,
2015; Scholtz et al., 2015). Evaluation of various operating gases shows that the reactive oxygen spe-
cies such as O, O3, OH, NO, and NO2 are the primary factors responsible for inactivating microbes
(Deng et al., 2006). The addition of oxygen gas in the operating gas mixture enhances the efficiency
of plasma treatment. The first experimental work on oxygen-based plasma was proposed in 1989
which reported lethal effect of oxygen gas-assisted plasma due to interference with biological matter.
Nelson and Berger (1989) also reported effectiveness of oxygen-based plasma which shows bacteri-
cidal effect against highly resistant bacteria species called B. subtilis and Clostridium sporogenes.
Niemira (2012) reported that use of nitrogen as a feed gas reduced the antimicrobial efficiency of
CP as compared to dry air while treating almonds inoculated with Salmonella and E. coli species.
15.6.2 Direct and Indirect treatment
Generally, it is assumed that the microbial inactivation efficiency of plasma was more in direct treat-
ment as compared to indirect treatment method. The possible reason could be that in direct treatment
sample is placed between the electrodes under plasma discharge leading to higher exposure of sample
to all generated reactive species. However, in case of indirect exposure, i.e., keeping the sample at
some distance from the discharge source, interaction between the reactive species and the sample is
comparatively reduced. This could be due to recombining nature of charged particles before reaching
to the sample, where only long-lived reactive species are reaching to the sample and causing lethal
effect (Laroussi 2009). Ziuzina et al. (2012) reported that direct plasma treatment was more effective
for inactivation of E. coli inoculated in liquid media inside a sealed package as compared to indirect
treatment. Fridman et al. (2007) also investigated the efficiency of atmospheric pressure plasma for
microbial inactivation by direct treatment. The results show that the inactivation was much faster
when bacterial cells come in contact with the charged particles of directly applied plasma discharge.
However, Surowsky et al. (2014) reported in their study that in liquid sample direct contact between
plasma and bacterial cells (i.e., Citrobacter freundii) is not required to achieve successful inactivation.
In case of liquid sample plasma treatment leads formation of highly reactive species, such as H2O2 and
hydroperoxyl radicals within the sample, therefore effectively inactivating microbes.
15.6.3 Other Factors
The level of interaction between microorganisms and the substrate, interference of substrate
with the treatment process such as absorption of active species or formation of new species or the
catalytic effect of substrate are some other factors affecting microbial inactivation efficiency of
plasma (in case of metal and ceramics). Lerouge et al. (2001) reported that substrate properties are
highly responsible for variation in microbial inactivation efficiency. Stratakos and Koidis (2015) also
explained that the change in substrate material, substrate type, and the characteristics of microor-
ganisms like microbial load, type, and their physiological states are most frequently observed inter-
related factors affecting the efficiency of plasma treatment. Song et al. (2009) investigated the effect
of substrate type on microbial inactivation efficiency of barrier discharge plasma. They inoculated
cheese and ham samples with cocktail of 3-strain of L. monocytogenes and exposed to barrier
discharge plasma. The treatment shows increase in microbial inactivation efficiency of plasma with
increasing treatment power (i.e., 75, 100, 125, and 150 W) and time (60, 90, and 120 s). The reduc-
tion in microbial count in treated cheese sample was found from 1.7 to 8 log cfu/g after 120 s of
treatment, whereas sliced ham showed only 0.25 to 1.73 log cfu/g reduction after same level of treat-
ment. They also reported that type of food material has strong association with plasma efficiency.
The microbial inactivation efficiency of plasma is moisture dependent which was observed higher
during the treatment of moist biological cell compared to lowest in dry cell (Dobrynin et al., 2009).
The interaction of reactive species of plasma with water molecules forms hydroxyl radicals which
are highly reactive leading to harmful effect on cells (Zou et al., 2004). The formation of hydroxyl
radicals is worth important as they are responsible for damaging 90% of DNA by forming hydration
layer around it.
The concentration of microbial load in the sample is negatively influencing the inactivation
process. Yong et al. (2014) reported reduction in inactivation efficiency of plasma treatment with
increasing concentration of E. coli count in inoculated raw chicken breast samples. Fernandez et al.
(2011) also reported decrease in microbial inactivation efficiency of cold atmospheric plasma with
increasing concentration S. typhimurium inoculated in polycarbonate membrane filters.
Treatment time, distance between the sample, and the discharge source and direction of treat-
ment are also interrelated factors affecting microbial inactivation efficiency of plasma. The increase
in treatment time significantly reduced microbial count in food. However, lipid oxidation and slight
colour darkness were observed with increasing treatment time of meat. The changes in texture
properties were relatively less with increasing treatment time and could be comparable to those of
control meat samples (Jayasena et al., 2015). Tappi et al. (2014) observed that in apple slices activ-
ity of polyphenol oxidase enzyme decreased linearly (up to 42%) with increasing treatment time.
Kim et al. (2013a) reported reduction of S. typhimurium count in inoculated chicken breast and
pork loin. The treatment of samples on both-side for 5 min at a distance of 20 mm significantly
reduced the S. typhimurium count, e.g., 0.66 log CFU/g and 1.33 log CFU/g respectively. Niemira
(2012) investigated the effect of CP on Salmonella and E. coli inoculated in almond samples using
distance (2, 4, and 6 cm) as a treatment variable. The result shows that keeping distance of 6 cm
between the sample and the discharge source resulted higher reduction, i.e., 1.34 log cfu/ml after
20 s of treatment. Suhem et al. (2013) optimized the power and distance of discharge source form
the sample to achieve maximum inhibition efficiency of plasma. They reported that the applica-
tion of plasma power of 40 W for 25 min is most effective for inactivation of A. flavous growth on
agar media. Bursac et al. (2016) reported that for successful treatment it is important to optimize
the variables like treatment time, volume of sample, and gas flow/min. They observed increase in
anthocyanin content from 21% to 35% in pomegranate juice when treated under optimum conditions
(i.e., treatment time 3 min, sample volume 5 cm, and gas flow rate 0.75 dm/min).
15.7 LIMItatIONS OF pLaSMa prOCESSING
Plasma is recognized for its valuable features including low operating temperature, less heat loss,
high surface modification and microbial inactivation efficiency, and less cost (Ruma et al., 2016).
Application of NTP or CP is advantageous, but it also has some drawbacks limiting its uses in food
processing industries. In some cases, it was observed that plasma treatment enhanced lipid oxidation,
colour reduction, changes in texture, and increase in acidity of food items. Thirumdas et al. (2014)
encountered 20% increase in peroxide value of peanut samples when treated at higher temperature
for longer time. They reported that the increase in peroxide value of treated peanut samples may be
due to reactive species of plasma capable in oxidizing lipid molecules. Instrumental colour (L*, a*, b*)
values of peanut samples were also reduced by the treatment. Misra et al. (2014a) observed reduction
of colour values of strawberries when treated with cold atmospheric plasma. Grabowski et al. (2016)
observed reduced intensity of aroma of almost all spices; however, intensity of colour in some spices
was also reduced by the application of low-pressure CP. Hertwig et al. (2015a) also reported similar
results in red paprika powder, where considerable reduction in redness was observed after remote
plasma treatment for 5 min or more. In case of spinach leaves wilting and discoloration was observed
after 5 min of treatment (Klockow and Keener 2009). Kim et al. (2015) reported a decrease in pH of
milk after 10 min of plasma treatment. Yong et al. (2015a) observed that treatment of sliced cheddar
cheese using DBD plasma significantly reduced flavour and overall acceptance as well as increased
off-flavour with increasing treatment time. As the plasma treatment is a surface phenomenon, it is
quite difficult to inactivate the endogenous enzymes intact in the tissue of whole fruit (Mir et al. 2015).
15.8 FUtUrE prOSpECtS OF pLaSMa prOCESSING
Plasma processing is an emerging alternative technology of other novel non-thermal techniques for
disinfecting the surface of fresh produce as well as packaging materials. It can be used for the treatment
of wide range of products in food processing industries; however, many aspects of plasma treatment in
food (i.e., chemical and nutritional changes) remain unexplored. It can play important role in agriculture
as the treatment reduces the germination time and enhancing the rate of seed germination. Most of the
equipment for plasma discharge are mainly designed and developed for biomedical field and limited to
laboratory-scale application. The issues like changes in food constituents mainly lipids, vitamins, etc.
by the treatment of CP still need to be solved. As the plasma discharge generates highly reactive species
that interact with the biological cell causing oxidative damage and breakdown of nucleus (DNA, RNA);
therefore, mutagenic properties of plasma in humans also need to be studied. Further studies also
needed to explore the safety aspects and cost benefits. Once these issues are solved, the technology will
gain wide popularity and adaptation in food industries for multiple applications. One such application
could be for the treatment of solid and liquid waste effluents of food industries.
rEFErENCES
Adler, H.J., Fischer, P., Heller, A., Jansen, I., Kuckling, D., and Komber, H. (1999). Trends in polymer
chemistry. Acta Polymerica. 50: 232–239.
Akishev, Y., Grushin, M., Dyatko, N., Kochetov, I., Napartovich, A., and Trushkin, N. (2008). Studies on cold
plasma polymer surface interaction by example of PP- and PET-films. Journal of Physics D: Applied
Physics. 41: 235203–235216.
Anthon, G.E., and Barrett, D.M. (2002). Kinetic parameters for the thermal inactivation of quality-related
enzymes in carrots and potatoes. Journal of Agricultural and Food Chemistry. 50: 4119–4125.
Ataeefard, M., Moradian, S., Mirabedini, M., Ebrahimi, M., and Asiaban, S. (2009). Investigating the effect
of power/time in the wettability of Ar and O2 gas plasma-treated low-density polyethylene. Progress in
Organic Coatings. 64: 482–488.
Baier, M., Foerster, J., Schnabel, U., Knorr, D., Ehlbeck, J., Herppich, W.B., and Schluter, O. (2013). Direct
non-thermal plasma treatment for the sanitation of fresh corn salad leaves: Evaluation of physical and
physiological effects and antimicrobial efficacy. Postharvest Biology and Technology. 84: 81–87.
Banik, I., Kim, K.S., Yun, Y.I., Kim, D.H., Ryu, C.M., and Park, C.E. (2002). Inhibition of aging in plasma-
treated high-density polyethylene. Journal of Adhesion Science and Technology. 16: 1155–1169.
Banu, M.S. (2012). Cold plasma as a novel food processing technology. International Journal of Emerging
Trends in Engineering and Developments. 4: 803–818.
Basaran, P., Basaran-Akgul, N., and Oksuz, L. (2008). Elimination of Aspergillus parasiticus from nut surface
with low pressure cold plasma (LPCP) treatment. Food Microbiology. 25: 626–632.
Bermudez-Aguireet, D., Wemlinger, E., Pedrow, P., Barbosa-Canovas, G., and Garcia-Perez, M. (2013). Effect
of atmospheric pressure cold plasma (APCP) on the activation of Escherichia coli in fresh produce.
Food Control. 34: 149–157.
Bie, P., Pu, H., Zhang, B., Su, J., Chen, L., and Li, X. (2015). Structural characteristics and rheological proper-
ties of plasma-treated starch. Innovative Food Science and Emerging Technologies. 34: 196–204.
Bonizzoni, G., and Vassollo, E. (2002). Plasma physics and technology: Industrial applications. Vacuum. 64:
327–336.
Bora, B., Bhuyan, H., Favre, M., Wyndham, E., and Chuaqui, H. (2011). Diagnostic of capacitively coupled low
pressure radio frequency plasma: An approach through electrical discharge characteristic. International
Journal of Applied Physics and Mathematics. 1: 124–128.
Bubler, S., Ehlbeck, J., and Schluter, O.K. (2017). Pre-drying of plant related tissue using plasma processed
air: Impact on enzyme activity and quality attributes of cut apple and potato. Innovative Food Science
and Emerging Technologies. 40: 78–86.
Buckenhuskes, H.J., and Rendlen, M. (2004). Hygienic problems of phytogenic raw materials for food produc-
tion with special emphasis to herbs and spices. Food Science and Biotechnology. 13: 262–268.
Bursac, K.D., Putnik, P., Dragovic-Uzelac, V., Pedisic, S., Rezek Jambrak, A., and Herceg, Z. (2016). Effects
of cold atmospheric gas phase plasma on anthocyanins and color in pomegranate juice. Food Chemistry.
190: 317–323.
Butscher, D., Zimmermann, D., Schuppler, M., and Von-Rohr, P.R. (2016). Plasma inactivation of bacterial
endospores on wheat grains and polymeric model substrates in a dielectric barrier discharge. Food
Control. 60: 636–645.
Cal, M.P., and Schluep, M. (2014). Destruction of benzene with non-thermal plasma in dielectric barrier dis-
charge reactors. Environmental Progress. 20(3): 151–156.
Chalkha, A., Despenes, C., Janulyte, A., Zerega, Y., Andre, J., Brkic, B., and Taylor, S. (2015). A dc glow
discharge as a source of electrons for a portable mass spectrometer: characterization of the electron cur-
rent intensity and electron kinetic energy distribution. Plasma Sources Science and Technology. 24(1):
015001. doi:10.1088/0963–0252/24/1/015001
Chen, H.H., Chang, H.C., Chen, Y.K., Hung, C.L., Lin, S.Y., and Chen, Y.S. (2016). An improved process for
high nutrition of germinated brown rice production: Low-pressure plasma. Food Chemistry. 191: 120–127.
Chen, H.H., Chen, Y.K., and Chang, H.C. (2012). Evaluation of physico-chemical properties of plasma treated
brown rice. Food Chemistry. 135: 74–79.
Chiad, B.T., Al-Zubaydi, T.L., Khalaf, M.K., and Khudar, A.I. (2010). Characterization of low pressure
plasma-DC glow discharges. Indian Journal of Pure and Applied Physics. 48: 723–730.
Critzer, F.J., Kelly-Wintenberg, K., South, S.L., and Golden, D.A. (2007). Atmospheric plasma inactivation of
food borne pathogens on fresh produce surfaces. Journal of food Protection. 70: 2290–2296.
D’Agostino, R.P., Favia, P., Oehr, C., and Wetheimer, M.R. (2005). Polymer surface modification with mono-
functional groups of different type and density. Plasma Processes and Polymer. 2: 7–15.
Deng, S., Ruan, R., Mok, C.K., Huang, G., Lin, X., and Chen, P. (2007). Inactivation of Escherichia coli on
almonds using nonthermal plasma. Journal of Food Science. 72: 62–66.
Deng, X.T., Shi, J.J., and Kong, M.G. (2006). Physical mechanisms of inactivation of Bacillus subtilis spores
using cold atmospheric plasmas. IEEE Transactions on Plasma Science. 34: 1310–1316.
Dhayal, M., Lee, S., and Park, S. (2006). Using low pressure plasma for Carthamus tinctorium L. Seed surface
modification. Vacuum. 80: 499–506.
Dirks, B.P., Dobrynin, D., Fridman, G., and Mukhin, Y. (2012). Treatment of raw poultry with non-thermal
dielectric barrier discharge plasma to reduce Campylobacter jejuni and Salmonella enterica. Journal of
Food Protection. 75: 22–28.
Dixon, D., and Meenan, B.J. (2012). Atmospheric dielectric barrier discharge treatments of polyethylene, poly-
propylene, polystyrene and poly (ethylene terephthalate) for enhanced adhesion. Journal of Adhesion
Science and Technology. 26: 2325–2337.
Dobrin, D., Magureanu, M., Mandache, N.B., and Ionita, M.D. (2015). The effect of non-thermal plasma treat-
ment on wheat germination and early growth. Innovative Food Science and Emerging Technologies.
29: 255–260.
Dobrynin, D., Fridman, G., Friedman, G., and Fridman, A. (2009). Physical and biological mecha-
nisms of direct plasma interaction with living tissue. New Journal of Physics. 11: 115020–115046.
doi:10.1088/1367-2630/11/11/115020.
El-Aziz, M.F.A., Mahmoud, E.A., and Elaragi, G.M. (2014). Non thermal plasma for control of the Indian
meal moth, Plodiainterpunctella (Lepidoptera: Pyralidae). Journal of Stored Products Research. 59:
215–221.
Elez-Martinez, P., Aguilo-Aguayo, I., and Martin-Belloso, O. (2006). Inactivation of orange juice peroxidase
by high intensity pulsed electric fields as influenced by process parameters. Journal of the Science of
Food and Agriculture. 86: 71–81.
Faisal, A.Q.D., and Dawood, M.O. (2014). Design and construction of argon dc glow discharge plasma using
Al target. Journal of KUFA – Physics. 6(2): 1–6.
FDA. (2013). Draft Risk Profile: Pathogens and Filth in Spices 1–213. Report of the Center for Food Safety
and Applied Nutrition, Food and Drug Administration, Department of Health and Human Services.
Fernandez, A., Shearer, N., Wilson, D.R., and Thompson, A. (2011). Effect of microbial loading on the
efficiency of cold atmospheric gas plasma inactivation of Salmonella enteric serovar Typhimurium.
International Journal of Food Mircrobiology. doi:10.1016/j.ijfoodmicro.2011.02.038.
Filatova, I., Azharonoki, V., Kadyrov, M., Beljavsky, V., Gvozdov, A., Shik, A., and Antonuk, A. (2011). The
effect of plasma treatment of seeds of some grain and legumes on their sowing quality and productivity.
Romanian Journal of Physics. 56: 139–143.
Fridman, A. (2008). Plasma Chemistry. Cambridge University Press, New York.
Fridman, G., Brooks, A.D., Balasubramanian, M., Fridman, A., Gutsol, A., Vasilets, V.N., Ayan, H., and
Friedman, G. (2007). Comparison of direct and indirect effects of non-thermal atmospheric-pressure
plasma on bacteria. Plasma Processes and Polymers. 4: 370–375.
Garofulic, I.E., Jambrak, A.R., Milosevic, S., Dragovic-Uzelac, V., Zoric, Z., and Herceg, Z. (2015). The effect
of gas phase plasma treatment on the anthocyanin and phenolic acid content of sour cherry Marasca
(Prunus cerasus var. Marasca) juice. LWT–Food Science and Technology. 62: 894–900.
Gaunt, L.F., Beggs, C.B., and Georghiou, G.E. (2006). Bactericidal action of the reactive species produced
by gas-discharge non-thermal plasma at atmospheric pressure: A review. IEEE Transactions on Plasma
Science. 34: 1257–1269.
Gieraltowski, L., Julian, E., Pringle, J., Macdonald, K., Quilliam, D., Marsden-Haug, N., Saathoff-Huber, L.,
Vonstein, D., Kissler, B., Parish, M., Elder, D., Howad-King, V., Besser, J., Sodha, S., Loharikar,
A., Dalton, S., Williams, I., and Barton Behravesh, C. (2013). Nationwide outbreak of Salmonella
Montevideo infections associated with contaminated imported black and red pepper: Ware house mem-
bership cards provide critical clues to identify the source. Epidemiological Infection. 141: 1244–1252.
Giner, J., Gimeno, V., Barbosa-Canovas, G.V., and Martin, O. (2001). Effects of pulsed electric field process-
ing on apple and pear polyphenoloxidases. Food Science and Technology International. 7: 339–345.
Gotoh, K., Yasukawa, A., and Taniguchi, K. (2011). Water contact angles on poly (ethylene terephthalate) film
exposed to atmospheric pressure plasma. Journal of Adhesion Science and Technology. 25: 307–322.
Grabowski, M., Strzelczak, A., and Dabrowski, W. (2014). Low pressure cold plasma as an alternative method
for black pepper sterilization. Journal of Life Sciences. 8: 931–939.
Grabowski, M., Wojcik, M., and Dabrowski, W. (2016). Effect of low-pressure cold plasma on sensory quality
and polyphenols content of spices. Global Scholastic Research Journal of Multidisciplinary. 2: 1–14.
Guo, J., Huang, K., and Wang, J. (2015). Bactericidal effects of various non thermal plasma agents and the
influence of experimental conditions in microbial inactivation: A review. Food Control. 50: 482–490.
Gurol, C., Ekinci, F.Y., Aslan, N., and Korachi, M. (2012). Low temperature plasma for decontamination of
E. coli in milk. International Journal of Food Microbiology. 157: 1–5.
Guzel-Seydim, Z.B., Greene, A.K., and Seydim, A.C. (2004). Use of ozone in the food industry. LWT-Food
Han, X., Cantrell, W.A., Escobar, E., and Ptasinsk, S. (2014). Plasmid DNA damage induced by helium atmo-
spheric pressure plasma jet. The European Physical Journal D. 10: 1–7. doi:10.1140/epjd/e2014-40753-y.
Hertwig, C., Reineke, K., Ehlbeck, J., Erdogdu, B., Rauh, C., and Schluter, O. (2015a). Impact of remote
plasma treatment on natural microbial load and quality parameters of selected herbs and spices. Journal
of Food Engineering. 167: 12–17. doi:10.1016/j.jfoodeng.2014.12.017.
Hertwig, C., Reineke, K., Ehlbeck, J., Erdogdu, B., Rauh, C., and Schluter, O. (2015b). Decontamination of whole
black pepper using different cold atmospheric pressure plasma applications. Food Control. 55: 221–229.
Hsu, C.M., Kao, P.K., Yang, Y.J., and Cheng, I.C. (2015). Rapid atmospheric pressure plasma jet processed
porous materials for energy harvesting and storage devices. Coatings. 5: 26–38.
Hury, S., Vidal, D.R., and Desor, F. (1988). A parametric study of the destruction efficiency of Bacillus spores
in low pressure oxygen-based plasmas. Letters in Applied Microbiology. 26: 417–421.
Jayasena, D.D., Kim, H.J., Yong, H.I., Park, S., Kim, K., Choe, W., and Jo, C. (2015). Flexible thin-layer
dielectric barrier discharge plasma treatment of pork butt and beef loin: Effects on pathogen inactivation
and meat-quality attributes. Food Microbiology. 46: 51–57.
Jiafeng, J., Xin, H., and Ling, L. (2014). Effect of cold plasma treatment on seed germination and growth of
wheat. Plasma Science and Technology. 16(1): 54–58.
Jie, S., and Yiping, Q. (2015). The effects of gas composition on the atmospheric pressure plasma jet modifica-
tion of polyethylene film. Plasma Science and Technology. 17: 402–408.
Junkar, I., Modic, M., and Mozeti, M. (2015). Modification of PET surface properties using extremely non-
equilibrium oxygen plasma. Open Chemistry. 13: 490–496.
Kaushik, N.K., Kim, Y.H., Han, Y.G., and Choi, E.H. (2013). Effect of jet plasma on T98G human brain cancer
cells. Current Applied Physics. 13: 178–180.
Keener, K.M., Jensen, J.L., Valdramidis, V.P., Byrne, E., Connolly, J., Mosnier, J.P., and Cullen, P.J. (2012).
Decontamination of Bacillus subtilis Spores in a sealed package using a Non-thermal Plasma system.
In Plasma for bio-decontamination, Medicine and Food Security. pp 445–455. Springer, Dordrecht. doi
10.1007/978-94-007-2852-3_34.
Kim, B., Yun, H., Jung, S., Jung, H., Choe, W., and Jo, C. (2011). Effect of atmospheric pressure plasma on inac-
tivation of pathogens inoculated onto bacon using two different gas compositions. Food Microbiology.
28(1): 9–13.
Kim, H.J., Yong, H.I., Park, S., Kim, K., Choe, W., and Jo, C. (2015). Microbial safety and quality attributes
of milk following treatment with atmospheric pressure encapsulated dielectric barrier discharge plasma.
Kim, H.J., Yong, H.I., Park, S., Kim, K., Bae, Y.S., Choe, W., Oh, M.H., and Jo, C. (2013a). Effect of inactivating
Salmonella Typhimurium in raw chicken breast and pork loin using an atmospheric pressure plasma jet.
Journal of Animal Science and Technology. 55: 545–549.
Kim, H.J., Yong, H.I., Park, S., Kim, K., Choe, W., and Jo, C. (2013b). Microbial safety and quality attributes
of milk following treatment with atmospheric pressure encapsulated dielectric barrier discharge plasma.
Kim, H.J., Yong, H.I., Park, S., Kim, K., Kim, T.H., Choe, W., Oh, M.H., and Jo, C. (2014b). Effect of
atmospheric pressure dielectric barrier discharge plasma on the biological activity of naringin. Food
Chemistry. 160: 241–245.
Kim, J.E., Lee, D.U., and Min, S.C. (2014a). Microbial decontamination of red pepper powder by cold plasma.
Food Microbiology. 38: 128–136.
Kim, M.C., Yang, S.H., Boo, J.H., and Han, J.G. (2003). Surface treatment of metals using an atmospheric
pressure plasma jet and their surface characteristics. Surface and Coatings Technology. 174: 839–844.
Klockow, P.A., and Keener, K.M. (2009). Safety and quality assessment of packaged spinach treated with a
novel ozone-generation system. LWT-Food Science and Technology. 42: 1047–1053.
Korachi, M., Ozen, F., Aslan, N., Vannini, L., Guerzoni, M.E., Gottardi, D., and Ekinci, F.Y. (2015). Biochemical
changes to milk following treatment by a novel, cold atmospheric plasma system. International Dairy
Journal. 42: 64–69.
Kostov, K.G., Honda, R.Y., Alves, L.M.S., and Kayama, M.E. (2009). Characteristics of dielectric barrier
discharge reactor for material treatment. Brazilian Journal of Physics. 39: 2322–2325.
Kudra, T., and Majumdar, A.S. (2009). Advanced Drying Technologies. CRC Press, Boca Raton, 438p.
Laimer, J., and Stori, H. (2006). Glow discharges observed in capacitive radio-frequency atmospheric-pressure
plasma jets. Plasma Processes and Polymers. 10: 573–586.
Langmuir, I. (1928). Oscillations in ionized gases. Proceedings of the National Academy of Science of the
United States of America. 14: 627–637. http://www.jstor.org/stable/85270.
Laroussi, M. (2009). Low-temperature plasma for medicine? Plasma Science. IEEE Transactions on Plasma
Science. 37: 714–725.
Lee, H.J., Jung, H., Choe, W., and Ham, J.S. (2011). Inactivation of Listeria monocytogenes on agar and pro-
cessed meat surfaces by atmospheric pressure plasma jets. Food Microbiology. 28: 1468–1471.
Lee, H.J., Song, H.P., Jung, H., Choe, W., Ham, J.S., Lee, J.H., and Jo, C. (2012). Effect of atmospheric pres-
sure plasma jet on inactivation of Listeria Monocytogenes, quality and genotoxicity of cooked egg white
and yolk. Korean Journal of Food Science Analysis. 32: 561–570.
Lee, K.P.K., Ju, W.T., and Lee, Y. (2006). Sterilization of bacteria, yeast and bacterial endospores by atmo-
spheric-pressure cold plasma using helium and oxygen. Journal of Microbiology. 44: 269–75.
Lerouge, S., Wertheimer, M.R., and Yahia, L.H. (2001). Plasma sterilization: A review of parameters, mecha-
nisms and limitations. Plasmas and Polymers. 6: 175–188.
Lii, C.Y., Liao, C.D., Stobinski, L., and Tomasik, P. (2002). Behaviour of granular starches in low-pressure
glow plasma. Carbohydrate Polymers. 49: 499–507.
Lin, Y.H., and Adomaitis, R.A. (2001). Simulation and model reduction methods for an RF plasma glow discharge.
Journal of Computational Physics. 171: 731–752.
Loretz, M., Stephan, R., and Zweifel, C. (2010). Antimicrobial activity of decontamination treatments for
poultry carcasses: A literature survey. Food Control. 21: 791–804.
McKee, L.H. (1995). Microbial contamination of spices and herbs: A review. LWT-Food Science and
Technology. 28: 1–11.
Medard, N., Soutif, J.C., Poncin-Epaillard, F. (2002). CO2, H2O, and CO2/H2O plasma chemistry for poly-
ethylene surface modification. Langmuir. 18: 2246–2253.
Menashi, W.P. (1968). Treatment of surfaces. US Patent Pub No.: 3,383,163.
Mir, S.A., Bosco, S.J.D., Shah, M.A., Mir, M.M., and Sunooj, K.V. (2015). Variety difference in quality charac-
teristics, antioxidant properties and mineral composition of brown rice. Journal of Food Measurement
and Characterization. 10: 177–184. doi:10.1007/s11694-015-9291-y.
Mir, S.A., Shah, M.A., and Mir, M.M. (2016). Understanding the role of plasma technology in food industry.
Food Bioprocess Technology. 9(5): 734–750. doi10.1007/s11947-016-1699-9.
Mirabedini, S.M., Arabi, H., Salem, A., and Asiaban, S. (2007). Effect of low-pressure O2 and Ar plasma
treatment on the wettability and morphology of biaxial-oriented polypropylene (BOPP) film. Progress
in Organic Coatings. 60: 105–111.
Misra, N.N., Kaur, S., Tiwari, B.K., Kaur, A., Singh, N., and Cullen, P.J. (2015). Atmospheric pressure cold
plasma (ACP) treatment of wheat flour. Food Hydrocolloids. 44: 115–121.
Misra, N.N., Keener, K.M., Mosnier, J.P., Bourke, P., and Cullen, P.J. (2014b). In-package atmospheric pressure
cold plasma treatment on quality of cherry tomatoes. Journal of Bioscience and Bioengineering. 118:
177–182. doi:10.1016/j.jbiosc.2014.02.005.
Misra, N.N., Patil, S., Moiseev, T., Bourke, P., Mosnier, J.P., Keener, K.M., and Cullen, P.J. (2014a). In-package
atmospheric pressure cold plasma treatment of strawberries. Journal of Food Engineering. 125: 131–138.
Mogul, R., Bolshakov, A.A., Chan, S.L., and Stevens, R.M. (2003). Impact of low-temperature plasma on
Deinococcus radiodurans and biomolecules. Biotechnology Progress. 19: 776–783.
Moisan, M., Barbeau, J., Moreau, S., Pelletier, J., Tabrizian, M., and Yahia L.H. (2001). Low-temperature ster-
ilization using gas plasmas: A review of the experiments and an analysis of the inactivation mechanism.
International Journal of Pharmaceuticals. 226: 1–21.
Montenegro, J., Ruan, R., Ma, H., and Chen, P. (2002). Inactivation of E-coli O157: H7 using a pulsed non-
thermal plasma system. Journal of Food Science. 67: 646–648.
Montie, T.C., Kelly-Wintenberg, K., and Roth, J.R. (2002). An overview of research using the one atmo-
sphere uniform glow discharge plasma (OAUGDP) for sterilization of surfaces and materials. IEEE
Transactions on Plasma Science. 28: 41–50.
Moreau, M., Feuilloley, M., Veron, W., Meylheuc, T., Chevalier, S., Brisset, J.L., and Orange, N. (2007).
Gliding arc discharge in the potato pathogen Erwiniacarotovora subsp. atroseptica: Mechanism of lethal
action and effect on the membrane associated molecules. Applied Environmental Microbiology. 73:
5904–5910.
Muranyi, P., Wunderlich, J., and Heise, M. (2007). Sterilization efficiency of a cascaded dielectric barrier
discharge. Journal of Applied Microbiology. 103: 1535–1544.
Napartovich, A.P. (2001). Overview of atmospheric pressure discharges producing nonthermal plasma.
Plasma Process and Polymers. 6: 1–14.
Nehra, V., Kumar, A., and Dwivedi, H. (2008). Atmospheric non-thermal plasma sources. International
Journal of Engineering. 2: 53–68.
Nelson, C.L., and Berger, T.J. (1989). Inactivation of microorganisms by oxygen gas plasma. Current
Microbiology. 18: 275–276. doi:10.1007/BF01570305.
Niemira, B.A. (2012). Cold plasma reduction of Salmonella and Escherichia coli O157: H7 on almonds using
ambient pressure gases. Journal of Food Science. 77: 171–175. doi:10.1111/j.1750-3841.2011.02594.x.
Niemira, B.A., and Sites, J. (2008). Cold plasma inactivates Salmonella Stanley and Escherichia coli O157:
H7 inoculated on golden delicious apples. Journal of Food Protection. 71: 1357–1365.
Noriega, E., Sharma, G., Laca, A., and Diaz, M. (2011). Cold atmospheric gas plasma disinfection of chicken
meat and chicken skin contaminated with Listeria innocua. Food Microbiology. 28: 1293–1300.
Oh, Y.A., Roh, S.H., and Min, S.C. (2016). Cold plasma treatments for improvement of the applicability of
defatted soybean meal-based edible film in food packaging. Food Hydrocolloids. 58: 150–159.
Pandiyaraj, K.N., Selvarajan, V., Deshmukh, R.R., and Bousmina, M. (2008). The effect of glow discharge
plasma on the surface properties of poly (ethylene terephthalate) (PET) film. Surface & Coatings
Technology. 202: 4218–4226.
Pankaj, S.K., Bueno-Ferrero, C., Mishra, N.N. Milosavljevic, V., O’Donnell, C.P., Bourke, P., Keener, K., and
Cullen, P.J. (2014). Application of cold plasma technology in food packaging. Trends Food Science and
Pankaj, S.K., Bueno-Ferrer, C., Misra, N.N., O’Neill, L., Tiwari, B.K., Bourke, P., and Cullen, P.J. (2015). Dielectric
barrier discharge atmospheric air plasma treatment of high amylose corn starch films. LWT-Food Science
and Technology. doi:10.1016/j.l.wt.2015.04.027.
Pankaj, S.K., Misra, N.N., and Cullen, P.J. (2013). Kinetics of tomato peroxidase inactivation by atmospheric
pressure cold plasma based on dielectric barrier discharge. Innovative Food Science & Emerging
Technologies. 19: 153–157.
Pashkuleva, I., Azevedo, H.S., and Reis, R.L. (2008). Surface structural investigation of starch-based bioma-
terials. Macromolecular Biosciences. 8: 210–219.
Pashkuleva, I., Marques, A.P., Vaz, F., and Reis, R.L. (2005). Surface modification of starch based blends
using potassium permanganate-nitric acid system and its effect on the adhesion and proliferation of
osteoblast-like cells. Journal of Material Science-Material and Medicine. 16: 81–92.
Pasquali, F., Stratakos, A.C., Koidis, A., Berardinelli, A., Cevoli, C., Ragni, L., and Trevisani, M. (2016).
Atmospheric cold plasma process for vegetable leaf decontamination: A feasibility study on radicchio
(red chicory, Cichorium intybus L.). Food Control. 60: 552–559. doi:10.1016/j.foodcont.2015.08.043.
Patra, S., Anjum, S., Ray, A.R., and Gupta, B. (2015). Effect of CO2 plasma exposure on physico-chemical
properties of porous polycaprolactone scaffold. Polymer Bulletin. doi:10.1007/s00289-015-1582-2.
Pelletier, J. (1992). Sterilization by plasma processing. Aggressologie. 33: 457–477.
Percival, M. (1996). Antioxidants. Clinical Nutrition Insights. 1: 1–6.
Perez, E., and Lares, M. (2006). Chemical composition, mineral profile, and functional properties of canna
(canna edulis) and arrowroot (Maranta spp.) starches. Plant Foods for Human Nutrition. 60: 113–116.
Perni, S., Liu, D.W., Sharma, G., and Kong, M.G. (2008a). Cold atmospheric plasma decontamination of the
pericarps of fruit. Journal of Food Protection. 71: 302–308.
Perni, S., Sharma, G., and Kong, M.G. (2008b). Cold atmospheric plasma disinfection of cut fruit surfaces
contaminated with migrating microorganisms. Journal of Food Protection. 71: 1619–1625.
Perni, S., Sharma, G., and Kong, M.G. (2008c). Cold atmospheric plasma disinfection of cut fruit surfaces
contaminated with migrating microorganism. Journal of Food Protection. 71: 1619–1625.
Poutanen, K., Sozer, N., and Della Valle, G. (2014). How can technology help to deliver more of grain in cereal
food for a healthy diet? Journal of Cereal Science. 59: 327–336.
Quan, C.Z., Xiang, Y.Z., Qing, X.G., Li, H.L., Lin, H.Y., Lin, L., Hai, M., Wei, H.X., and Kudryavtsev, A.A.
(2015). Pulsed microwave driven argon plasma jet with distinctive plume patterns resonantly excited by
surface plasma on polaritons. Chinese Physics B. 24: 1–11.
Ragni, L., Berardinelly, A., Vnnini, L., Montanari, C., Sirri, F., Guerzoni, M.E., and Guarneri, A. (2010).
Non-thermal atmospheric gas plasma device for surface decontamination of shell eggs. Journal of Food
Engineering. 100: 125–132.
Ramazzina, I., Berardinelli, A., Rizzi, F., Tappi, S., Ragni, L., Sacchetti, G., and Rocculi, P. (2015). Effect of
cold plasma treatment on physic-chemical parameters and antioxidant activity of minimally processed
kiwifruit. Postharvest Biology and Technology. 107: 55–65.
Rauf, S., and Kushnerb, M.J. (1998). The effect of radio frequency plasma processing reactor circuitry on
plasma characteristics. Journal of Applied Physics. 83: 5087–5094.
Ruma, Habib, M.A., and Basha, R.H. (2016). A survey of non-thermal plasma and their generation methods.
International Journal of Renewable Energy and Environmental Engineering. 4, ISSN 2348-0157.
Sampedro, F., Rodrigo, M., Martinez, A., Rodrigo, D., and Barbosa-Canovas, G. (2005). Quality and safety aspects
of PEF application in milk and milk products. Critical Reviews in Food Science and Nutrition. 45: 25–47.
Sarangapani, C., Devi, Y., Thirundas, R., Annapure, U.S., and Deshmukh, R.R. (2015). Effect of low-pressure
plasma on physico-chemical properties of parboiled rice. LWT-Food Science and Technology. 63: 452–460.
Sarangapani, C., O’Toole, G., Cullen, P.J., and Bourke, P. (2017). Atmospheric cold plasma dissipation
efficiency of agrochemicals on blueberries. Innovative Food Science and Emerging Technologies.
doi:10.1016/j.ifset.2017.02.012.
Schneider, J., Akbar, M.I., Dutroncy, J., Kiesler, D., Leins, M., and Schulz, A. (2009). Silicon oxide barrier
coatings deposited on polymer materials for applications in food packaging industry. Plasma Processes
and Polymers. 6: S700–S704. doi:10.1002/ppap.200931702.
Schneider, J., Baumgartner, K. M., Feichtinger, J., Kruger, J., Muranyi, P, Schulz, A., Walker, M., Wunderlich, J.,
and Schumacher, U. (2005). Investigation of the practicability of low-pressure microwave plasmas in
the sterilization of food packaging materials at industrial level. Surface and Coatings Technology. 200:
962–966.
Scholtz, V., Pazlarova, J., Souskova, H., Khun, J., and Julak, J. (2015). Nonthermal plasma: A tool for decon-
tamination and disinfection. Biotechnology Advances. 33: 1108–1119.
Schwabedissen, A., Łacinski, P., Chen, X., and Engemann, J. (2007). Plasma label: A new method to disinfect goods
inside a closed package using dielectric barrier discharges. Contribution to Plasma Physics. 47: 551–558.
Schweiggert, U., Carle, R., and Schieber, A. (2007). Conventional and alternative processes for spice production:
A review. Trends in Food Science and Technology. 18: 260–268.
Segat, A., Misra, N.N., Cullen, P.J., and Innocente, N. (2015). Atmospheric pressure cold plasma (ACP) treat-
ment of whey protein isolate model solution. Innovative Food Science and Emerging Technologies. 29:
247–254.
Selcuk, M., Oksuz, L., and Basaran, P. (2008). Decontamination of grains and legumes infected with Aspergillus
spp. and penicillium spp. B y cold plasma treatment. Bioresource Technology. 99: 5104–5109.
Shainsky, N., Dobrynin, D., Ercan, U., Joshi, S.G., Ji, H., Brooks, A., Fridman, G., Cho, Y., Fridman, A.,
and Friedman, G. (2012). Plasma acid: Water treated by dielectric barrier discharge. Plasma Processes
Polymer. 10: 1–6.
Shakila, B.M., Sasikala, P., Dhanapal, A., Kavitha, V., Yazhini, G., and Rajamani, L. (2012). Cold plasma
as a novel food processing technology. International Journal of Emerging Trends in Engineering and
Development. 4: 803–818.
Shao, C., Wang, D., Tang, X., Zhao, L., and Li, Y. (2013). Stimulating effects of magnetized arc plasma of dif-
ferent intensities on the germination of old spinach seeds. Mathematical and Computer Modelling. 58:
814–818.
Shi, X.M., Zhang, G.J., Wu, X.L., and Li, Y.X. (2011). Effect of low-temperature plasma on microorganism
inactivation and quality of freshly squeezed orange juice. IEEE Transactions on Plasma Science. 39:
1591–1597.
Slepicka, P., Slepickova Kasalkova, N., Stranska, E., Bacakova, L., and Svorcik, V. (2013). Surface characteriza-
tion of plasma treated polymers for applications as biocompatible carriers. Express Polymer Letters. 7:
535–545.
Slepicka, P., Vasina, A., Kolská, Z., Luxbacher, T., Malinsky, P., Macková, A., and Švorcík, V. (2010). Argon
plasma irradiation of polypropylene. Beam Interact. Nuclear Atoms and Methods in Physics Research B.
268: 2111–2114.
Smulders, E.H.W.M., Heesch, B.E.J.M.V., and Paasen, S.S.V.B.V. (1998). Pulsed power corona discharges for
air pollution control. IEEE Transactions on Plasma Science. 26(5): 1476–1484.
Soloshenko, I.A. (2000). Sterilization of medical products in low-pressure glow discharges. Plasma Physics
Reports. 26: 792–800.
Song, H.P.B., Kim, J.H., Choe, S., Jung, S.Y., Moon, W., and Choe, C.J. (2009). Evaluation of atmospheric
pressure plasma to improve the safety of sliced cheese and ham inoculated by 3-strain cocktail Listeria
monocytogenes. Food Microbiology. 26: 432–436.
Starzyk, F., Lii, C.Y., and Tomasik, P. (2001). Light absorption, transmission and scattering in potato starch
granule. Polish Journal of Food and Nutrition Sciences. 10: 27–34.
Stratakos, A.C., and Koidis, A. (2015). Suitability, efficiency and microbiological safety of novel physical
technologies for the processing of ready-to-eat meats and pumpable products. International Journal of
Food Science and Technology. 50: 1283–1302.
Suhem, K., Matan, N., Nisoa, M., and Matan, N. (2013). Inhibition of Aspergillus flavus on agar media
and brown rice cereal bars using cold atmospheric plasma treatment. International Journal of Food
Microbiology. 161: 107–111.
Suresh, D., Manjunatha, H., and Srinivasan, K. (2007). Effect of heat processing of spices on the concen-
tration of their bioactive principles: Turmeric (Curcuma longa), red pepper (Capsicum annuum) and
black pepper (Piper nigrum). Journal of Food Composition and Analysis. 20: 346–351. doi:10.1016/j.
jfca.2006.10.002.
Surowsky, B., Fischer, A., Schlueter, O., and Knorr, D. (2013). Cold plasma effects on enzyme activity in a
model food system. Innovative Food Science and Emerging Technology. 19: 146–152.
Surowsky, B., Frohling, A., Gottschalk, N., Schluter, O., and Knorr, D. (2014). Impact of cold plasma on
Citrobacter freundii in apple juice: Inactivation kinetics and mechanisms. International Journal of
Szymanowskia, H., Kaczmareka, M., and Gazicki-Lipmana, M. (2005). New biodegradable material based on
RF plasma modified starch. Surface Coating Technology. 200: 539–543.
Tanino, M., Xilu, W., Takashima, K., Katsura, S., and Mizuno, A. (2005). Sterilization using dielectric barrier
discharge at atmospheric pressure. IEEE. 2: 784–788. doi:10.1109/IAS.2005.1518421.
Tappi, S., Berardinelli, A., Ragni, L., Dalla Rosa, M., Guarnieri, A., and Rocculi, P. (2014). Atmospheric
gas plasma treatment of fresh cut apples. Innovative Food Science and Emerging Technologies. 21:
114–122.
Tendero, C., Tixier, C., Tristant, P., Desmaison, J., and Leprince, P. (2006). Atmospheric pressure plasma:
A review. Spectrochimica Acta Part B: Atomic Spectroscopy. 61: 2–30.
Terrier, O., Essere, B., Yver, M., Barthelemy, M., Bouscambert-Duchamp, M., Kurtz, P., Van Mechelen, D.,
Morfin, F., Billaud, G., Ferraris, O., Lina, B., Rosa-Calatrava, M., and Moules, V. (2009). Cold oxy-
gen plasma technology efficiency against different airborne respiratory viruses. Journal of Clinical
Virology. doi:10.1016/j.jcv.2009.03.017.
Thakur, B.R., and Nelson, P.E. (1998). High-pressure processing and preservation of food. Food Review
International. 14: 427–447.
Thirumdas, R., Sarangapani, C., and Annapure, U.S. (2014). Cold plasma: A novel non-thermal technology for
food processing. Food Biophysics. 10: 1–11.
Tiwari, B.K., O’Donnell, C.P., and Cullen, P.J. (2009). Effect of non thermal processing technologies on the
anthocyanin content of fruit juices. Trends in Food Science & Technology. 20: 137–145.
Tsuji, A., Yasaka, Y., Kang, S.Y., Morimoto, T., and Sawada, I. (2008). A practical simulation scheme for slot-
excited microwave plasma reactor equipped with dual shower plate. Thin Solid Films. 516: 4368–4373.
Van de Veen, H.B., Xie, H., and Esveld, E. (2014). Inactivation of chemical and heat-resistant spores of
Bacillus and Geobacillus by nitrogen cold atmospheric plasma evokes distinct changes in morphology
and integrity of spores. Food Microbiology. 45: 26–33. doi:10.1016/j.fm.2014.03.018.
Vesel, A., Kovac, J., Primc, G., Junkar, I., and Mozetic, M. (2016). Effect of H2S plasma treatment on the
surface modification of a polyethylene terephthalate surface. Materials. doi:10.3390/ma9020095.
Vesel, A., and Mozetic, M. (2012). Surface modification and aging of PMMA polymer by oxygen plasma
treatment. Vacuum. 86: 634–637.
Vleugels, M., Sharma, G., Deng, X.T., and Greenacre, E. (2005). Atmospheric plasma inactivation of biofilm-
forming bacteria for food safety control. IEEE Transactions on Plasma Science. 33: 824–828.
Volin, J.C., Denes, F.S., Young, R.A., and Park, S.M.T. (2000). Modification of seed germination performance
through cold plasma chemistry technology. Crop Science. 40: 1706–1718.
Wanga, T.C., Lua, N., Li, J., and Wu, Y. (2010). Degradation of pentachlorophenol in soil by pulsed corona
discharge plasma. Journal of Hazard. 180: 436–441.
Wissel, S.A., Zwicker, A., Ross, J., and Gershman, S. (2013). The use of DC glow discharges as undergraduate
educational tools. American Journal of Physics. 81: 663–669.
Yong, H.I., Kim, H.J., Park, S., Alahakoon, A.U., Kim, K., Choe, W., and Jo, C. (2015a). Evaluation of pathogen
inactivation on sliced cheese induced by encapsulated atmospheric pressure dielectric barrier discharge
plasma. Food Microbiology. 46: 46–50. doi:10.1016/j.fm.2014.07.010.
Yong, H.I., Kim, H.J., Park, S., Choe, W., Oh, M.W., and Jo, C. (2014). Evaluation of the treatment of both sides
of raw chicken breasts with an atmospheric pressure plasma jet for the inactivation of Escherichia coli.
Food Borne Pathogens and Disease. 11: 652–657.
Yong, H.I., Kim, H.J., Park, S., Kim, K., Choe, W., Yoo, S.J., and Jo, C. (2015b). Pathogen inactivation and
quality changes in sliced cheddar cheese treated using flexible thin-layer dielectric barrier discharge
plasma. Food Research International. 69: 57–63.
Yoon, Y.H., and Ryu, K.H. (2007). Atmospheric plasma surface treatment equipment. News Information
Chemical Engineering. 25: 268–271.
Yun, H., Kim, B., Jung, S., Kruk, Z.A., Kim, D.B., Choe, W., and Jo, C. (2010). Inactivation of Listeria mono-
cytogenes inoculated on disposable plastic tray, aluminium foil and paper cup by atmospheric pressure
plasma. Food Control. 21: 1182–1186.
Zhang, L., Lu, Z., Lu, F., and Bie, X. (2006). Effect of γ irradiation on quality-maintaining of fresh-cut lettuce.
Zhang, M., Oh, J.Y., Cisneros-Zevallos, L., and Akbulut, M. (2013). Bactericidal effects of nonthermal low-
pressure oxygen plasma on S. typhimurium LT2 attached to fresh produce surfaces. Journal of Food
Engineering. 119: 425–432.
Zhong, K., Wu, J., Wang, Z., Chen, F., Liao, X., and Hu, X. (2007). Inactivation kinetics and secondary structural
change of PEF-treated POD and PPO. Food Chemistry. 100: 115–123.
Ziuzina, D., Patil, S., Cullen, P.J., Keener, K.M., and Bourke, P. (2012). Atmospheric cold plasma inactivation of
Escherichia coli in liquid media inside a sealed package. Journal of Applied Microbiology. 114: 778–787.
Zou, J.J., Liu, C.J., and Eliasson, B. (2004). Modification of starch by glow discharge plasma. Carbohydrate
Polymers. 55: 23–26.
ChaptEr 16
Electron Beam processing of Foods
Shima Shayanfar and Suresh D. Pillai
CONtENtS
16.1 Introduction ....................................................................................................................... 315

16.2 Electron Beam Technology Parameters ............................................................................ 316
16.3 Different Applications of eBeam in Food Processing ...................................................... 318
16.4 Bacterial Response to eBeam............................................................................................ 319
16.5 Fruits and Vegetables ........................................................................................................ 319
16.6 Cereal Products ................................................................................................................. 320
16.7 Meat and Poultry Products ................................................................................................ 321
16.8 Egg and Milk Products ..................................................................................................... 321
16.9 Spices ................................................................................................................................ 322
16.10 Space Food ........................................................................................................................ 322
16.11 Regulations and Standards ................................................................................................ 323
References ...................................................................................................................................... 325
16.1 INtrODUCtION
Electron beam (eBeam) irradiation is one of the main three principal ionizing radiation tech-
niques (gamma, X-ray, eBeam) that are available to myriads of different industries including the
food industry. The working principle is based on speeding up electrons (from commercial elec-
tricity) that are generated off a cathode in a vacuum environment. An electron gun consisting of
a cathode, grid, and anode generates and accelerates the beam. A magnet focuses the generated
beam of electrons in order to control the pattern in which the beam leaves the gun. Considering
the source of energy, speed, cost, safety, etc., electron beam accelerators are becoming preferred to
cobalt-60-based gamma ray food irradiation technology (Clemmons et al., 2015). One of the main
advantages of eBeam technology that differentiates it from gamma irradiation is that it is a “switch-
on/switch-off” technology, meaning it can be switched on when needed and off when not needed.
Ionizing radiation results in DNA and RNA strand breakage in microorganisms and thus the growth
in spoilage or pathogenic microorganisms is hampered.
The earliest type of irradiation introduced to the food industry was gamma irradiation in 1960
as an alternative method to canning (Roberts, 2014). Although the original development of electron
beam linear accelerators took place during the 1930s, it took some time for the technology to be
315
table 16.1 the history of Food Irradiation

Year Milestone
1905 First patent on application of irradiation in foodstuff by Appleby J and banks AJ (uK)
1921 use of X-ray for inactivation of trichinae in raw pork by schwartz b (us)
1930 French patent on inactivation of bacteria in packaged food using X-rays
1956 Lewis proposes the first electron beam accelerator
1957 First commercial application of ebeam for spices in Germany
1958 us FDA evaluates food irradiation as a food additive and not as a physical process.
1958–1959 ussr approves potato and grain irradiation
1960 Canada approves potato irradiation
1963–1964 us FDA approves irradiated bacon, wheat, wheat flour, and potatoes
1964–1967 us FDA approves flexible packaging for food containers during irradiation
1966 First international symposium of Food irradiation in Karlsruhe, Germany
1976 Joint FAo/iAeA/WHo expert Committee approves several irradiated foods and recommends food
irradiation as a physical process
1980 Joint FAo/iAeA/WHo expert Committee approves all foods to be irradiated up to 10 kGy
1983–1984 Codex Alimentarius sets up standard for food irradiation
1986 us FDA approves fresh fruits and vegetables irradiation up to 1 kGy
1979–1990 Assisting developing countries by training and demonstrating food irradiation facilitates,
Wageningen, the netherlands
1994 Fsis indicates the radiation doses for red meat products.
2000–2004 electron beam-based food irradiation facilities established in the us
2003 national Center for electron beam research established at texas A&M university
2014 electron beam technology for phytosanitary treatment of imported fruits in the us
A review of the published data indicates that the quantity of irradiated foods in the world is approximately
405,000 tons (table 16.2).
commercialized (Lung et al., 2015). The technology has developed over time to become a safe and
efficient food processing technology (Farkas et al., 2014; Pillai and Shayanfar, 2015). The history of
food irradiation is summarized in Table 16.1.
16.2 ELECtrON BEaM tEChNOLOGY paraMEtErS
When adopting electron beam technology for food irradiation, one has to have a very clear idea
of the primary goal of the application. Is it for food pasteurization? Is it for decontaminating or
sterilizing spices and ingredients? Is it for phytosanitary applications? The answers to these ques-
tions will drive the decisions in terms of the equipment specifications, the product handling, control
systems, the shielding, etc., which in turn will dictate the financial and technological sustainability of
the proposed eBeam project. Global status of irradiation and challenges in use of eBeam technology
are given in Tables 16.2 and 16.3, respectively.
Phytosanitary irradiation relies on the use of ionizing radiation such as eBeam for disinfecting
fruits and vegetables from insects and pests. Since insects and pests are more sensitive than bac-
terial cells to ionizing radiation, the doses used in phytosanitary treatment are in the range of
100–1000 Gy and are significantly lower than the doses used for food pasteurization, which are
in the range of 1–10 kGy (in most parts of the world). For disinfecting or sterilizing spices and
ingredients, doses in the range of 15–30 kGy are often employed. The dose is the amount of energy
absorbed per unit area (measured in Grays or kilograys).
eLeCtron beAM ProCessinG oF FooDs 317
table 16.2 the Global Status of Food Irradiation in 2005

Food application percentage (%) amount (ton)
spices and dry Disinfestation 46 186,000
vegetables
Garlic and potato sprout inhibition 22 88,000
Grains and fruits Disinfestation 20 82,000
Meat and fish Disinfestation 8 32,000
others Mushroom, honey, etc. 4 17,000
total 100 405,000
Source: Kume, t. et al., Radiat. Phys. Chem., 78, 222–226, 2009.

Unique features associated with eBeam have made it a promising alternative to
the other non-thermal technologies in addressing safety, quality, and sustainability
in food processing (Table 16.3).
table 16.3 Benefits and Contemporary Challenges associated

with application of eBeam technology
Benefits
• non-thermal technology
• no loss in nutrients, flavours, or appearance of food
• Applicable on packaged products (avoids cross contamination)
• no addition of chemicals to achieve product safety and quality
• Low carbon footprint
• Low energy requirements
• Very high processing speeds
• not dependent on radioactive materials
• Adjustable for varying doses and different applications
• Continuous process
• Precise doses
Contemporary Challenges
• Widespread confusion and misinformation about the technology
• Limited vendors providing the technology
• technology not widely available due to costs of adoption
• Lower penetration depth when compared to gamma irradiation
In terms of the equipment and the technology specifications, the linear accelerator energy
(measured as million electron volts, MeV) and linear accelerator power (measured in kilowatts, kW)
are extremely critical. The linear particle accelerator (often shortened as linac) energy dictates the
penetrating ability of the electrons. The penetration of eBeam into food packages depends on the
areal density (g/cm2) of the package. Areal density defined as mass per unit area is critically impor-
tant when assessing whether eBeam irradiation is possible. When employing a 10 MeV eBeam lin-
ear accelerator, only products with areal density less than 8.3 g/cm2 can be processed with eBeam
irradiation. Reconfiguration of the packaging dimensions to satisfy the areal density constraints
is the most common option employed to adopt eBeam processing. The linac’s power dictates the
processing speed of the eBeam system. The throughput of an eBeam system is calculated by linac
power/minimum dose = kg/sec. The minimum dose is established by understanding the sensitivity
of the target organism(s) to the eBeam process.
In addition to adhering to the applicable doses allowed doses by the regulatory agencies, accu-
rate information about the sensitivity of the target microorganism is essential. The sensitivity of
microorganisms to ionizing radiation such as eBeam is expressed as D10-value, which is the dose
required in order to reduce the initial population of microorganisms by one log or 90%. Different
microorganisms, different species, and even different strains and isolates of a species can have
different D10 values because of their inherent physiological differences. Fungal spores and bac-
terial endospore states are significantly more resistant than their vegetative states. Another fac-
tor that enhances the efficiency of ionizing radiation such as eBeam technology is the moisture
content of the product. The higher the moisture content or water activity in the food product, the
more efficient the technology is. The reason is creation of high-reactive species arising from the
radiolytic splitting of water that can cause significant damage to the microbial DNA. The results
of some earlier research claim that at doses higher than 10 kGy, lipid oxidation should be of con-
cern; however, such high doses are rarely applied in the majority of fresh foods. Other factors such
as temperature, water pH, and chemical composition of food can affect the irradiation efficiency
(Sommers, 2012; Roberts, 2014).
16.3 DIFFErENt appLICatIONS OF eBEaM IN FOOD prOCESSING
The primary metric used in food irradiation unit is dose (measured in Gy [Grays] or kGy
[1 kGy = 1000 Gy]). The different applications of the technology are defined by the target doses as
shown in the table below (Table 16.4). Table 16.4 is the list of food items that is approved for irradia-
tion in the US by the FDA. The eBeam doses applied in food processing can be broadly categorized
into three different dose ranges:
1. Low-dose treatment (<1 kGy); applied for phytosanitary, insect disinfestation, delaying the maturity
in fruits, preventing germination (e.g., potato, onion, ginger, etc.)
2. Intermediate-dose treatment (1–10 kGy), food pasteurization and extension of shelf life of foods
3. High-dose treatment (10–44 kGy), sterilization or disinfection of spices and ingredients
table 16.4 FDa-approved applications of Irradiation technology for Food processing

application Dose (kGy)
Control of Trichinella spiralis in pork carcasses 0.3–1
inhibition of fresh food growth and maturation <1
Control of Salmonella in fresh shell eggs <3.0
Control of food-borne pathogens and extension of shelf life in fresh iceberg lettuce and fresh spinach <4.0
Control of food-borne pathogens in fresh meat products <4.5
Control of food-borne pathogens in fresh poultry products <4.5
Control of Vibrio bacteria and other pathogens in fresh or frozen molluscan shellfish <5.5
Control of food-borne pathogens and extension of shelf life of chilled, frozen, raw, cooked, <6.0
partially cooked, or dried crustaceans
Control of food-borne pathogens in frozen meat and poultry products <7.0
Control of microbial pathogens on seeds for sprouting <8.0
Microbial disinfection of dry or dehydrated enzyme preparations <10
Microbial disinfection of dry or dehydrated aromatic vegetables, herbs, and spices <30
sterilization of space food >44
Source: FDA, CFr-Code of Federal regulations title 21. Part 179 – irradiation in the production, processing and
handling of food. Available at: http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfCFr/CFrsearch.
cfm?CFrPart=179andshowFr=1, 2016.
16.4 BaCtErIaL rESpONSE tO eBEaM
Various microorganisms are inactivated by eBeam by direct damage to their DNA by the highly
energetic electrons or indirectly by the radiolytic products formed when the highly energetic elec-
trons interact with aqueous solutions in the food. Exposure of aqueous streams to eBeam irradiation
causes extensive ionization reactions and the generation of free radicals and electrons as follows
(Eq 16.1):
e − + H 2O → [ 2.6] eaq + [ 0.55] H + [ 2.7] H 3O + + [ 0.7] H 2O2 + [ 2.6] HO + [ 0.55] H 2 (16.1)

− • •
Where, the values in brackets represent “G values” (number of species produced by 0.1 MeV of
energy absorbed), and H•, e−, and HO • are highly reactive species, while H3O+ is the hydrated
proton. The technology is paradigm shifting, because it simultaneously creates both reduction and
oxidation processes without the addition of any chemicals. These powerful oxidation-reduction
reactions occur almost instantaneously and are therefore characterized as an advanced oxidation-
reduction process (AORP). The eBeam technology is the most cost-effective approach of generating
free radicals (Tahergorabi et al., 2012; Lung et al., 2015). Therefore, as was mentioned earlier,
increasing moisture content of the products enhances the efficacy of the irradiation.
Different microorganisms have different levels of sensitivity to specific doses of eBeam.
Some species of microorganisms (e.g., Deinococcus radiodurans) can survive the common low
doses applied for food decontamination and might even resume growth (Patterson et al., 1993).
Therefore, in order to completely inactivate such eBeam resistant species higher doses of eBeam
are required. However, it should be considered that this higher required dose should not exceed
the doses permitted by FDA or it can affect the organoleptic properties of the food (Tahergorabi
et al., 2012). Therefore, when processing foods with eBeam technology, both pathogen sensitivity
to eBeam and retention of food/sensory quality parameters should be considered. On the other
hand, sub-lethal damages arising from a synergistically applied food processing technology can
also increase the sensitivity of the target organisms to eBeam doses. The eBeam processing can
be complementarily used along with other contemporary food processing technologies (Pillai and
Shayanfar, 2015).
16.5 FrUItS aND VEGEtaBLES
The health benefits of fresh fruits and vegetables are widely known. The World Health
Organization (WHO) also encourages the daily intake of at least 400 g of fruit and vegetable per day
for the prevention of various diseases, such as heart disease, cancer, diabetes, and obesity (Callejón
et al., 2015). Fresh produce is mainly consumed for the naturally occurring phytochemicals, which
are highly susceptible to heat treatments. However, they might also carry pathogenic microorgan-
isms that jeopardize the lives of consumers. Fresh produce is the major cause of many of the food-
borne outbreaks (CSPI, 2015). Therefore, the non-thermal status of ionizing radiation makes it
an exciting technique in order to ensure both safety and quality in fresh produce. The majority of
the studies on irradiation of fresh produce were performed using gamma irradiation and thus the
number of studies performed with eBeam on fresh produce is not as exhaustive as that of gamma
irradiation. The first marketed irradiated strawberries were treated with gamma in January 1992
(Marcotte, 1992) and the success encouraged the irradiation of strawberries. According to FDA
regulations for irradiation of fresh produce (except for fresh iceberg lettuce and spinach), no fresh
fruits or vegetable can be irradiated with doses higher than 1 kGy (FDA, 2016). Although this dose
has been mainly suggested for extending shelf life through delayed maturation it has other benefits
including microbial inactivation. Some of the benefits of eBeam processing on fresh produce are:
• Non-thermal inactivation of both spoilage and pathogenic microorganisms

• Phytosanitary treatment of agricultural products to meet global phytosanitary regulations
• Extending shelf-life without altering the sensory attributes or consumer preferences
• Preventing sprouting
• Preventing post-harvest losses in grains and pulses from insects and pests
• Decontamination of food processing wastes
• Non-thermal sterilization of packaging materials for aseptic packaging
Application of only 2.3 kGy eBeam in cabbage resulted in more than 4-log reduction in the popula-
tion of bioburden microorganisms (Grasso et al., 2011). Furthermore, they showed that increasing
the dose to 4.0 kGy could decrease the population of E. coli K-12 by about 7 logs. In another study
application of 0.7 kGy of eBeam could reduce 5 logs of the test organism in baby spinach leaves
packed under modified atmosphere packaging (MAP) (Gomes et al., 2011). A dose of 1 kGy of
eBeam could reduce 4.5 logs of the population of a cocktail of big six E. coli strains in fresh straw-
berry puree (Shayanfar et al., 2016). Application of 7 kGy of eBeam on tomato, cantaloupe, and
lettuce seeds could reduce Salmonella population by 3–5 logs (Trinetta et al., 2011). Espinosa et al.
(2012) showed that eBeam at 4 kGy could significantly reduce rotavirus and poliovirus in iceberg
lettuce and spinach about 2.5 and 3 log units, respectively.
16.6 CErEaL prODUCtS
Cereal grains are susceptible to a variety of damages caused by insects, rodents, and microor-
ganisms throughout their journey from farm to post-harvest practices. Considering the processing
cycle of cereal and grains there is no efficient kill step to ensure food safety. Recent pathogenic
E. coli outbreaks associated with consumption of cookie dough and cake mix (CDC, 2009, 2016)
have highlighted the necessity of introducing a kill step that not only ensures food safety but also
does not affect the functionality of major components of grains such as gluten.
An irradiation treatment up to 5 kGy has been proved to improve cereal quality without
impairing the physiochemical properties (Rao et al., 1978; Kim et al., 2012). However, the applica-
bility of this dose in the US is questionable because the FDA has limited the upper dose limit for
flour to be 500 Gy (FDA, 2016).
The effect of gamma irradiation on different grains has been extensively studied (Rao et al.,
1978; MacArthur et al., 1983; Köksel et al., 1998; Kanemaru et al., 2005; Kim et al., 2012);
however, there is still limited information about application of eBeam on cereal (Sitton et al.,
1995; Hu et al., 2010; Kim et al., 2012). Application of low doses of gamma (0.0, 0.5, 1.0, and
2.0 kGy) on wheat flour has indicated that there is no influence on protein content of the wheat
flour. However, a decrease in falling number and an increase in water absorption of the grains have
been observed (MacArthur et al., 1983; Kanemaru et al., 2005; Singer et al., 2006). Additionally,
weaker dough was obtained from irradiated grains, whereas, there was no such effect noticed in
the dough made of irradiated flour. The mentioned doses of gamma did not affect dough develop-
ment (MacArthur et al., 1983; Kanemaru et al., 2005). Increasing the irradiation dose to 3 kGy
increased the farinograph absorption while the dough development time and stability decreased
(MacArthur et al., 1983). However, in the study conducted by Singer et al. (2006), increasing the
dose up to 2 kGy did not affect the dough development and the gas retention in the baked bread as
well as colour of the grains. On the other hand, higher doses of up to 10 kGy are known to leave
important effects on cereal grain quality including weakening of dough mixture properties and
protein changes.
Application of doses up to 4.6 kGy of eBeam did not have negative effect on wheat quality except
for reduced seed germination and seedling vigour at higher doses at 10.2 kGy, while it reduced Tilletia
tritici infection of wheat from 75.5% to 0.08% (Sitton et al., 1995). In another study by Hu et al. (2010)
application of eBeam up to 4.4 kGy caused a slight decrease in gelatinization temperature because of the
starch granules damage increased from 5.33% to 7.38% as the eBeam dose increased from 0 to 4.4 kGy.
16.7 MEat aND pOULtrY prODUCtS
An extensive body of research on the irradiation of fresh meat and poultry products has occurred
over the past 40 years. A considerable amount of information on all aspects of meat and poultry
irradiation has been developed over this period (Giddings and Marcotte, 1991; Lee et al., 1996).
About 18 million pounds of ground beef is being commercially irradiated in the US for retail and
commercial sales. There is no exact figure available on to what percent this amount is processed
with eBeam, but it is assumed the eBeam is applied on 50% of this volume.
The efficacy of irradiation on inoculated poultry meat and ground beef with different patho-
genic microorganisms including Salmonella typhimurium, Pseudomonas putida, Escherichia coli,
Streptococcus faecalis, Staphylococcus aureus, Listeria monocytogenes, and Lactobacillus spp.
(Patterson, 1988; Giddings and Marcotte, 1991; Thayer, 1995) has been established. In general, a
dose of 2.5–5.0 kGy does not alter any of the organoleptic properties of the meat or poultry products
and can also extend the shelf life at cold temperature from 6 to 14 days (Mulder, 1984). The results
of research also suggest that in general vacuum or carbon dioxide atmosphere during irradiation
offers the most lethal effect on pathogens (Patterson, 1988). Most of the pathogens can be inacti-
vated with a single dose of <3.0 kGy; however, enteric viruses and the endospores of Clostridium
and Bacillus are highly resistant to ionizing radiation (Thayer, 1995). Therefore, different hurdles
have been proposed to enhance the efficiency of irradiation (Vas, 1981).
It is essential to bear in mind eBeam processing by no means should be considered a killing
step in meat processing but to be acknowledged as the final critical control point in a validated
hazard analysis and critical control point (HACCP) system. The main advantage of eBeam in meat
processing is the fact that it can be applied as a post-packaging process, which limits the incidences
of cross contamination of the meat to pathogens such as E. coli and Salmonella spp.
16.8 EGG aND MILK prODUCtS
The increasing number of food-borne outbreaks associated with Salmonella contamination of

eggs has sparked the interest of irradiating egg products. Various studies have been performed to
determine the proper irradiation dose for egg products and to evaluate the potential effects of dif-
ferent does on functional properties of egg products (Brooks et al., 1959; Katusin-Razem et al.,
1989; Narvaiz et al., 1992). The results of this research show that a dosage of 2–3 kGy can inacti-
vate Salmonella without any major changes in the sensory properties (Katusin-Razem et al., 1989;
Narvaiz et al., 1992). The eBeam-treated raw yolk at 2.5 kGy showed improved emulsion capacity
over the untreated ones after 7 days of storage and retained more soluble protein; however, eBeam-
treated egg yolk was less bright. No significant difference in low-density lipoprotein (LDL) was
observed between eBeam-treated and untreated egg yolk samples. eBeam irradiation did not cause
any significant physical, chemical, or functional changes in egg yolk at 2.5 kGy (Huang et al., 1997).
Consumption of raw milk has gained a lot of popularity recently in belief that more nutrients can be
obtained from raw milk (Oliver et al., 2009). However, a considerable number of food-borne outbreaks
associated with consumption of raw milk or products made out of it are reported (Claeys et al., 2013;
CDC, 2014; Jayarao et al., 2006). In order to serve this emergent need there is a need for application of a
non-thermal pasteurization technique such as eBeam processing to ensure safety without compromising
the quality attribute. However, there is no standard suggested by FDA with regard to irradiation of milk
or milk products (FDA, 2016). There are limited studies conducted to understand the feasibility of
application of eBeam as a cold pasteurization method without affecting the quality (Ward et al., 2017).
The results of application of 1–2 kGy of eBeam on raw milk did not indicate any significant changes
in lactose, vitamin B12, or calcium concentrations and although there was about 31.6% decrease in
vitamin B2 concentration, the content still met USDA nutritional guidelines. The GC-olfactory analy-
sis also did not result in the development of a wide variety of off-odours. These results support the fact
the eBeam can be considered as a promising processing technique to address major concerns regard-
ing the pathogens in raw milk without affecting the quality attributes (Ward et al., 2017).
16.9 SpICES
Spices are both economically and functionally high-value commodities for both flavour and
colour. At the same time, spices can be contaminated with various bacteria, moulds, and pathogens
originating in the soil as well as poor hygienic conditions that occur during post-harvest process-
ing. Therefore, it is essential to consider a decontamination method that addresses the microbio-
logical safety without compromising the aroma and/or flavour of spices. The most common spice
contaminants are Bacillus cereus and Clostridium perfringens. These can cause a variety of food-
borne illnesses. Among spices imported into the US, Salmonella is the predominant pathogen found
in spices imported from Asia. Furthermore, the presence of the secondary metabolites of certain
moulds that contaminate spices to lead to aflatoxin contamination (Jeswal and Kumar, 2015).
Irradiation with eBeam can be considered the most effective method in ensuring safety in
spices. Application of eBeam technology achieves the reduction of food-borne pathogens, opportu-
nistic pathogens, and spoilage microbes without compromising the initial flavours, colours, aromas,
and antioxidant properties in the spices. The amount of spices irradiated commercially has been
estimated to be about 100,000 tons worldwide in 2002 (Kitazuru et al., 2004).
In a study by Kume et al. (1989) the radiosensitivity of toxigenic Aspergilli isolated from spices
against gamma irradiation was investigated. The D10 values and induction doses were reported to be
267–293 Gy and 75–155 Gy in wet conditions, respectively. A dose of 8 kGy was suggested for destruc-
tion of Aspergilli according to the calculated values. However, in an attempt to reduce ochratoxin A
(OTA) and aflatoxins B1, B2, G1, and G2 (AFB1, AFB2, AFG1, and AFG2) in black pepper, Jalili et al.
(2010) applied a range of 0–60 kGy of gamma irradiation on mycotoxin concentration ranging from 10
to 100 ng g−1. The maximum mycotoxin reduction was found at 60 kGy of about 52%, 43%, 24%, 40%,
and 36% for OTA, AFB1, AFB2, AFG1, and AFG2, respectively. The study results clearly indicate that
even 60 kGy of gamma rays is inefficient for complete destruction of ochratoxin and aflatoxins. In fact,
even at a dose of 30 kGy, which is the maximum permitted level by the FDA, mycotoxin reduction is
less than 30%. The bacterial counts of commercially available spices vary between 102 and 103 CFU/g;
however, application of 7.5–10 kGy of irradiation was adequate to sterilize pepper, cardamom, and nut-
meg (Sharma et al., 1984). In another study evaluating the efficiency of irradiation on decontamination
of Saffron the D-value for the mould, bacterium, and yeast were reported to be 0.82, 0.86, and 2.69 kGy,
respectively, indicating how yeasts are resistant to ionizing radiation (Ghoddusi and Glatz, 2004).
16.10 SpaCE FOOD
Thermal sterilization of foods has extended the shelf life of space food to 3–5 years. However,
the amount of energy required to achieve all the safety parameters and the potential quality determi-
nation has encouraged irradiation of space food (Catauro and Perchonok, 2012). The FDA oversees
the regulations of space food irradiation and obligates a minimum dose of 44 kGy. There are further
studies going on at Texas A&M University to attenuate this dose to lower doses where both sensory
quality and microbiological safety are addressed (Bhatia et al., 2016).
16.11 rEGULatIONS aND StaNDarDS
Despite the fact that eBeam technology is extensively deployed in over 60 countries there are
some regional regulations that limit the type of the foods that can be irradiated. The safety of food
irradiation under 10 kGy has been proven by a Joint Expert Committee on Food Irradiation consist-
ing of WHO, IAEA, and FAO (JECFI, 1981). There is a specific list of the food items that can be
irradiated in the USA and EU. In the USA, FDA bans the irradiation of cooked foods, dairy prod-
ucts, and juice (FDA, 2016). The regulations governing food irradiation are relatively more relaxed
in countries such as India (Tables 16.5 and 16.6).
In China, cooked meats along with pollen, fruits, honey, beans, grains, spices, and dehy-
drated vegetables are allowed to be irradiated (Chen et al., 2012). In Brazil, for instance,
there is no limit set over the maximum irradiation dose permitted to be applied in various
food items as long as the sensory attributes are not changed (Niemira and Deschenes, 2005).
table 16.5 Indian regulations Governing Ionization Irradiation Dose Limits and applications for
Different Classes of Foods
Dose Limits (kGy)
Class Food type application Minimum Maximum
1 bulbs, stem and root tubers, and inhibit sprouting 0.02 0.2
rhizomes
2 Fresh fruits and vegetables (other than Delay ripening 0.2 1.0
class 1) insect disinfestation 0.2 1.0
shelf-life extension 1.0 2.5
Quarantine 0.25 1.0
3 Cereals and their milled products, pulses insect disinfestation 0.25 1.0
and their milled products, nuts, oil bioburden reduction 1.5 5.0
seeds, dried fruits, and their products
4 Fish, aquaculture, seafood, and their Pathogen elimination 1.0 7.0
products (fresh or frozen) and shelf-life extension 1.0 3.0
crustaceans
Control of protozoan parasites 0.3 2.0
5 Meat and meat products including poultry Pathogen elimination 1.0 7.0
(fresh and frozen) and eggs shelf-life extension 1.0 3.0
Control of protozoan 0.3 2.0
pathogens
6 Dry vegetables, seasonings, spices, bioburden reduction 6.0 14.0
condiments, dry herbs and their insect disinfestation 0.3 1.0
products, tea, coffee, cocoa, and plant
products
7 Dried foods of animal origin and their Pathogen elimination 2.0 7.0
products Control of moulds 1.0 3.0
insect disinfestation 0.3 1.0
8 ethnic foods, military rations, space bioburden reduction 2.0 10.0
foods, ready-to-eat, ready-to-cook, and Quarantine application 0.25 1.0
minimally processed foods
sterilization 5.0 25.0
table 16.6 Indian regulations Governing Dose Limits for Ionizing radiation processing of Food-
related allied products
Dose Limits (kGy)

allied product application Minimum Maximum
Packaging materials for food and allied products bioburden reduction 5.0 10.0
Food additives bioburden reduction 5.0 10.0
Health foods, dietary supplements, and nutraceuticals bioburden reduction 5.0 10.0
table 16.7 European Union Countries with Current Food Irradiation programs
European Union Country Food Items being Irradiated

belgium Dehydrated blood, egg white, fish, shellfish, shrimp, frog legs, gum Arabic, herbs
and spices, poultry, rice meal, vegetables
Czech republic Herbs and spices
Germany Herbs and spices
estonia Herbs and spices
spain Herbs and spices
France Frog legs, gum Arabic, herbs and spices, poultry
Hungary Herbs and spices
netherlands egg whites, fish, shellfish, shrimp, frog legs, herbs and spices, poultry, dehydrated
products
Poland Herbs and spices
Source: Pillai, s.D., Chem. Eng. Prog., 112, 36–44, 2016.
The European Union approves food irradiation and food irradiation is currently ongoing in
the EU (Table 16.7).
Labelling of foods in the US is regulated by the FDA, which requires that the radura symbol
(Figure 16.1) has to be used along with the phrase, “treated with irradiation” or “treated by irra-
diation” or “irradiated for food safety.” Unlike the EU where even ingredients are irradiated, the
US does not require foods prepared from irradiated ingredients to be labelled. Irradiated foods
Figure 16.1 radura symbol.

(e.g., fruits treated for phytosanitary treatment) that are not packaged have to be displayed at the
point of sale with placards or banners or posters.
In addition to food irradiation regulations, there are a number of regulations governing the
irradiation facility. Since eBeam equipment is considered as radiation-producing device, there
are a number of specific regulations that govern radiation shielding, ozone removal, occupational
safety, etc. (Brown, 2015).
rEFErENCES
Bhatia, S.S., Wall, K.R., Kerth, C.R., and Pillai, S.D. (2016). Benchmarking the minimum electron beam
(eBeam) dose required for the sterilization of space foods. Radiation Physics and Chemistry. 143:72–78.
Brooks, J., Hannan, R.S., and Hobbs, B.C. (1959). Irradiation of eggs and egg products. International Journal
of Applied Radiation Isotopes. 6:149–154.
Brown, D. (2015). Integrating electron beam equipment into food processing facilities: Strategies and design
considerations. In: Electron Bean Pasteurization and Complementary Food Processing Technologies,
pp. 27–46. Pillai, S.D., and Shayanfar, S., Eds., Woodhead Publishing, Cambridge, UK.
Callejón, R.M., Rodríguez-Naranjo, M.I., Ubeda, C., Hornedo-Ortega, R., Garcia-Parrilla, M.C., and Troncoso,
A.M. (2015). Reported foodborne outbreaks due to fresh produce in the United States and European
Union: Trends and causes. Food Borne Pathogens and Disease. 12(1):32–38.
Catauro, P.M., and Perchonok, M.H. (2012). Assessment of the long-term stability of retort pouch foods to
support extended duration spaceflight. Journal of Food Science. 77:S29–S39.
CDC. (2009). Multistate outbreak of E. coli O157:H7 infections linked to eating raw refrigerated, prepackaged
cookie dough. Available at: https://www.cdc.gov/ecoli/2009/cookie-dough-6-30-2009.html.
CDC. (2014). Raw (unpasteurized) milk. Available at: http://www.cdc.gov/Features/RawMilk/index.html.
Accessed March 2017.
CDC. (2016). Multistate outbreak of Shiga toxin-producing Escherichia coli infections linked to flour.
Available at: https://www.cdc.gov/ecoli/2016/o121-06-16/.
Chen, S.Q., Jiang, K.X., Cao, B.S., Liu, Y.Q., Cai, X.F., Liu, X.L., Cao, Y., Liu, X.Y., and Shi, W.N. (2012).
Distribution of irradiated foods in China. Food Control. 28(2):237–239.
Claeys, W.L., Cardoen, S., Daube, G., Block, D.J., Dewettinck, K., Dierick, K., De Zutter, L. et al. (2013).
Raw or heated cow milk consumption: Review of risks and benefits. Food Control. 31:251–262.
Clemmons, H.E., Clemmons, E.J., and Brown, E.J. (2015). Electron beam processing technology for food
processing. In: Electron Bean Pasteurization and Complementary Food Processing, pp. 11–25. Pillai,
S.D., and Shayanfar, S., Eds., Woodhead Publishing, Cambridge, UK.
CSPI. (2015). Outbreak alert! 2015. A review of foodborne illness in the U.S. from 2004–2014. Center for Science
in the Public Interest. Available at: https://cspinet.org/sites/default/files/attachment/outbreak-alert-2015.pdf.
Espinosa, A.C., Jesudhasan, P., Arredondo, R., Cepeda, M., Mazari-Hiriart, M., Mena, K.D., and Pillai, S.D.
(2012). Quantifying the reduction in potential health risks by determining the sensitivity of poliovirus
type 1 chat strain and rotavirus SA-11 to electron beam irradiation of iceberg lettuce and spinach.
Applied and Environmental Microbiology. 78(4):988–93.
Farkas, J., Ehlermann, D.A.E., and Mohacsi-Farkas, C.S. (2014). Food irradiation. In: Encyclopedia of Food
Safety: Foods, Materials, Technologies and Risks, Vol. 3, pp. 178–186. Motarjemi, Y., Ed., Elsevier.
FDA. (2016). CFR-Code of Federal Regulations Title 21. Part 179 – Irradiation in the production, processing
and handling of food. Available at: http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfCFR/
CFRSearch.cfm?CFRPart=179andshowFR=1, accessed March 2017.
Ghoddusi, H.B., and Glatz, B. (2004). Decontamination of saffron (Crocus sativus L.) by electron beam
irradiation. Acta Horticulture. 650:339–344.
Giddings, G.G., and Marcotte, M. (1991). Poultry irradiation: For hygiene/safety and market‐life enhancement.
Food Reviews International. 7(3):259–282.
Gomes, C., Moreira, R.G., and Castell-Perez, E. (2011). Radiosensitization of Salmonella spp. and Listeria
spp. in ready-to-eat baby spinach leaves. Journal of Food Science. 76:141–148.
Grasso, E.M., Uribe-Rendon, R.M., and Lee, K. (2011). Inactivation of Escherichia coli inoculated onto
fresh-cut chopped cabbage using electron-beam processing. Journal of Food Protection. 74:115–118.
Hu, B., Huang, M., Yin, S., Zi, M., and Wen, Q. (2010). Effects of electron-beam irradiation on physicochemical
properties of starches separated from stored wheat. Starch. 63(3):121–127.
Huang, S., Herald, T.J., Mueller, D.D. (1997). Effect of electron beam irradiation on physical, physiochemical,
and functional properties of liquid egg yolk during frozen storage. Poultry Science. 76(11):1607–1615.
Jalili, M., Jinap, S., and Noranizan, A. (2010). Effect of gamma radiation on reduction of mycotoxins in black
pepper. Food Control. 21(10):1388–1393.
Jayarao, B.M., Donaldson, S.C., Straley, B.A., Sawant, A.A., Hedge, N.V., and Brown, J.L. (2006). A sur-
vey of foodborne pathogens in bulk tank milk and raw milk consumption among farm families in
Pennsylvania. Journal of Dairy Science. 89(7):2451–2458.
JECFI. (1981). Wholesomeness of Irradiated Food: Report of a Joint FAO/IAEA/WHO Expert Committee,
WHO Technical Report Series, 659, World Health Organization, Geneva, Switzerland.
Jeswal, P., and Kumar, D. (2015). Mycobiota and natural incidence of aflatoxins, ochratoxin, and
citrinin in Indian spices confirmed by LC-MS/MS. International Journal of Microbiology.
doi:10.1155/2015/242486.
Kanemaru, J., Tavares, D.T., Singer, C.S., Hilsenrath, F.C., Sabato S.F., Tadini, C.C. (2005). Influence of
gamma irradiation on rheological properties of wheat flour. Eurotherm Seminar 77 – Heat and Mass
Transfer in Food Processing, June 20–22, Parma, Italy.
Katusin-Razem, B., Razem, D., Matic, S., Mihokovic, V., Kostromin-Soos, N., and Milanovic, N. (1989).
Chemical and organoleptic properties of irradiated dried whole egg and egg yolk. Journal of Food
Protection. 52:781–786.
Kim, G.R., Akram, K., Ahn, J.J., and Kwon, J.H. (2012). Identification of gamma ray and electron-beam irra-
diated wheat after different processing treatments. Journal of Cereal Science. 56:347–351.
Kitazuru, E.R., Moreira, A.V.B., Mancini-Filho, M., Delincee, H., and Villavicencio, A.L.C.H. (2004). Effects
of irradiation on natural antioxidants of cinnamon (Cinnamomum zeylanicum N.). Radiation Physics
and Chemistry. 71:37–39.
Köksel, H., Sapirstein, H.D., Celik, S., and Bushuk, W. (1998). Effects of gamma-irradiation of wheat on glu-
ten proteins. Journal of Cereal Science. 28:243–250.
Kume, T., Furuta, M., Todoriki, S., Uenoyama, N., and Kobayashi, Y. (2009). Status of food irradiation in the
world. Radiation Physics and Chemistry. 78:222–226.
Kume, T., Ito, H., and Soedarman, H. (1989). Radiosensitivity of toxigenic Aspergillus isolated from spices
and destruction of aflatoxins by gamma-irradiation. International Journal of Radiation Applications
and Instrumentation. Part C. Radiation Physics and Chemistry. 34(6):973–978.
Lee, M., Sebranek, J.G., Olson, D.G., and Dickson, J.S. (1996). Irradiation and packaging of fresh meat and
poultry. Journal of Food Protection. 59(1):62–72.
Lung, H.M., Cheng, Y.C., Chang, Y.H., Huang, H.W., Yang, B.B., and Wang, C.Y. (2015). Microbial decon-
tamination of food by electron beam irradiation. Trends in Food Science and Technology. 44:66–78.
MacArthur, L.A., and D’Appolonia, B.L. (1983). Gamma radiation of wheat. I. Effects on dough and baking
properties. Cereal Chemistry. 60(6):456–460.
Marcotte, M. (1992). Irradiated strawberries enter the U.S. market. Food Technology. 46(5):80–86.
Mulder, R.W.A.W. (1984). Ionizing energy treatment of poultry. Food Technology Association of Australia.
36:418–420.
Narvaiz, P., Lescano, G., and Kairiyama, E. (1992). Physicochemical and sensory analyses on egg powder
irradiated to inactivate Salmonella and reduce microbial load. Journal of Food Safety. 12:263–282.
Niemira, B.A., and Deschenes, L. (2005). Ionizing radiation processing of fruits and fruit products. In:
Processing Fruits: Science and Technology, pp. 221–252. Barrett, D.M., Somogyi, L., and Ramaswamy,
H., Eds., CRC Press, Elsevier, Woodhead Publishing, Boca Raton.
Oliver, S.P., Boor, K.J., Murphy, S.C., and Murinda, S.E. (2009). Food safety hazards associated with con-
sumption of raw milk. Foodborne Pathogenic Disease. 6(7):793–806.
Patterson, M. (1988). Sensitivity of bacteria to irradiation on poultry meat under various atmospheres. Letters
in Applied Microbiology. 7(3):55–8.
Patterson, M.F., Damouglou, A.P., and Buick, R.K. (1993). Effect of irradiation on poultry meat and in phos-
phate-buffered saline. Letters in Applied Microbiology. 8:181–184.
Pillai, S.D. (2016). Introduction to electron-beam food irradiation. Chemical Engineering Progress.
112(11):36–44.
Pillai, S.D., and Shayanfar, S. (2015). Electron Bean Pasteurization and Complementary Food Processing
Technologies. Woodhead Publishing, Cambridge, UK.
Rao, V.S., Vakil, K.C., Bandyopadhyay, C., and Sreenivasan, A. (1978). Effect of gamma irradiation of wheat
on volatile flavor components of bread. Journal of Food Science. 43:68–71.
Roberts, P.B. (2014). Food irradiation is safe: Half a century of studies. Radiation Physics and Chemistry.
105:78–82.
Sharma, A., Ghanekar, A.S., and Padwal-Desai, S.R. (1984). Microbiological status and antifungal properties
of irradiated spices. Journal of Agricultural and Food Chemistry. 32(S):1061–1063.
Shayanfar, S., Mena, K.D., and Pillai, S.D. (2016). Quantifying the reduction in potential infection risks from
non-O157 Shiga toxin producing Escherichia coli in strawberries by low dose electron beam processing.
Food Control. doi:10.1016/j.foodcont.2016.04.05.
Singer, C.S., Sabato, S.F., and Tadini, C.C. (2006). Breadmaking properties of flour produced from irradiated
wheat. CIGR Section VI International Symposium on Future of Food Engineering, Warsaw, Poland,
April 26–28.
Sitton, J.W., Borsa, J., Schultz, T.R., and Maguire, J.D. (1995). Electron beam irradiation effects on wheat
quality, seed vigor, and viability and pathogenicity of teliospores of Tilletia controversa and T. tritici.
Plant Disease. 79(6):586–589.
Sommers, C.H. (2012). Microbial decontamination of food by irradiation. In: Microbial Decontamination
in the Food Industry, Novel Methods and Applications, pp. 322–343. Sommers, C.H., Ed., Woodhead
Publishing Series in Food Science, Technology and Nutrition.
Tahergorabi, R., Matak, K.E., and Jaczynski, J. (2012). Application of electron beam to inactivate Salmonella
in food: Recent developments. Food Research International. 45:685–694.
Thayer, D.W. (1995). Use of irradiation to kill enteric pathogens on meat and poultry. Journal of Food Safety.
15(2):181–192.
Trinetta, V., Vaidya, N., Linton, R., and Morgan, M. (2011). A comparative study on the effectiveness of chlo-
rine dioxide gas, ozone gas and e-beam irradiation treatments for inactivation of pathogens inoculated
onto tomato, cantaloupe and lettuce seeds. International Journal of Food Microbiology. 146:203–206.
Vas, K. (1981). Technological feasibility of combination treatments. Combination processes in food irradiation.
Proceedings of a Symposium, pp. 125–130, Colombo, Sri Lanka, November 24–28, International Atomic
Energy Agency, Vienna, Austria.
Ward, L.R., Kerth, C.R., and Pillai, S.D. (2017). Nutrient profiles and volatile odorous compounds
of raw milk after exposure to electron beam pasteurizing doses. Journal of Food Science.
doi:10.1111/1750-3841.13763.
ChaptEr 17
Combination of Non-thermal processes

and their hurdle Effect
Swati Sethi, Rahul Kumar Anurag, Yogesh Kumar, and O. P. Chauhan
CONtENtS
17.1 Introduction ........................................................................................................................ 330

17.2 Concept of Hurdle Effect .................................................................................................... 331
17.3 Non-thermal Processing Techniques in Combination with Conventional Hurdles ............ 336
17.3.1 High Hydrostatic Pressure in Combination with Conventional Preservation
Technologies ....................................................................................................... 336
17.3.1.1 Combination of High Hydrostatic Pressure with
Acidification (Low pH) .................................................................... 337
17.3.1.2 High Hydrostatic Pressure in Combination with Heat ....................... 338
17.3.1.3 High Hydrostatic Pressure in Combination with Antimicrobials ......340
17.3.2 Ultrasonication in Combination with Conventional Preservation Technologies ....341
17.3.2.1 Ultrasonication in Combination with Heat (Thermo-sonication) ...... 342
17.3.2.2 Ultrasonication in Combination with Antimicrobials ........................ 343
17.3.3 Continuous Ultraviolet Processing with Possible Hurdles .................................344
17.3.4 Pulsed Light Processing in Combination with Conventional Preservation
Hurdles ...............................................................................................................346
17.3.4.1 Pulsed Light Processing in Combination with Heat .......................... 347
17.3.4.2 Pulsed Light Processing in Combination with Antimicrobials.......... 347
17.3.5 Pulsed Electric Field in Combination with Conventional Preservation Hurdles ..... 348
17.3.5.1 Pulsed Electric Field in Combination with Heat ................................ 349
17.3.5.2 Pulsed Electric Field in Combination with Acidic pH ....................... 349
17.3.5.3 Pulsed Electric Field in Combination with Antimicrobial................. 350
17.3.6 Ozone in Combination with Conventional Preservation Technologies .............. 350
17.3.6.1 Ozone in Combination with Heat ....................................................... 351
17.3.6.2 Ozone in Combination with Antimicrobials ...................................... 351
17.3.7 Electron Beam Processing in Combination with Conventional
Preservation Hurdles ...................................................................................... 352
17.3.7.1 Electron Beam Processing in Combination with Antimicrobials ...... 352
329
17.4 Hurdle Approach in Combining Non-thermal Technologies ............................................. 352

17.4.1 High Hydrostatic Pressure and Ozone ............................................................... 353
17.4.2 High Hydrostatic Pressure and Microfiltration .................................................. 353
17.4.3 High Hydrostatic Pressure and Carbon dioxide ................................................. 353
17.4.4 High Hydrostatic Pressure and Gamma Irradiation........................................... 354
17.4.5 Ultrasonication and Supercritical Carbon Dioxide ............................................ 354
17.4.6 Ultrasonication and Osmotic Pressure (Osmo-sonication) ................................ 354
17.4.7 Ultrasonication and Ultraviolet Radiation (Photosonication) ............................ 355
17.4.8 Ultrasonication and Pulsed Light Processing .................................................... 355
17.4.9 Ultrasonication and Ionizing Radiation Processing ........................................... 356
17.4.10 Ultrasonication and Pulsed Electric Field Processing ....................................... 356
17.4.11 Ultrasonication and High Hydrostatic Pressure (Mano-sonication) .................. 356
17.4.12 Ultrasonication, Heat and High Hydrostatic Pressure
(Mano-thermo-sonication) ............................................................................... 357
17.4.13 Pulsed Electric Field and High-Pressure Carbon dioxide .................................. 357
17.4.14 Pulsed Electric Field and Ozone ........................................................................ 358
17.4.15 Pulsed Electric Field and Microfiltration ........................................................... 358
17.4.16 Pulsed Electric Field and High Hydrostatic Pressure ........................................ 358
17.4.17 High Hydrostatic Pressure and Ultraviolet Light Assisted Titanium dioxide
Photocatalysis (TiO2-UV photocatalysis) ........................................................... 359
17.4.18 Ozone and Ultrasonication .................................................................................360
17.5 Current Status in Hurdle Approach Towards Non-thermal Processing Technologies
at Laboratory and Commercial Level ................................................................................360
17.6 Future Scope in Hurdle Approach Towards Non-thermal Processing Technologies ......... 361
References ...................................................................................................................................... 362
17.1 INtrODUCtION
The knowledge of traditional or conventional thermal food processing has been utilized in
enhancing food safety for ages due to its safety profile, technological simplicity, commercial
viability, profitability, and consumer acceptability. Thermal processing technologies prevail as
an ideal approach in the food industry for preserving and extending the shelf life of foods.
Such technologies; however, involve application of high temperatures for a sufficiently long
time to destroy pathogenic as well as spoilage microorganisms, in production of shelf-stable
food products. This, in turn, induces irreversible changes to quantifiable factors, such as nutrient
degradation, and non-quantifiable factors, such as sensory attributes of food. Consequently, with
time- and market-driven demand, efforts have been made to improve the quality of these prod-
ucts by utilizing non-conventional thermal processing technologies such as microwave, infrared,
and ohmic heating which work on the principal of volumetric heating. Therefore, these technolo-
gies are able to maintain the nutritional quality to some extent due to shorter processing times.
However, in the present era, minimal processing has received considerable attention due to
consumer demand of fresh-like or natural food products, which have made the food processing
industry seek alternative or novel approaches to achieve desired food safety goals. To overcome
the drawbacks of thermal processing effects, non-thermal processing technologies, minimal
processing, and advanced food packaging techniques have evolved in the recent two decades in
food processing operations.
CoMbinAtion oF non-tHerMAL ProCesses AnD tHeir HurDLe eFFeCt 331
These non-thermal technologies work at ambient or near-ambient temperature, thereby

having potential to retain the desired quality in terms of nutritional and organoleptic quality
of food product. Non-thermal technologies have the prospective to provide high-quality,
fresh-like produce with considerable high shelf-life. The food industry is actively involved in
these technological developments and poised to adopt new alternatives that offer competitive
advantages. Current research trends are concentrated towards finding commercial applications
of these non-thermal processes. However, anti-microbiological capabilities, safety aspects,
technical difficulties, process efficiency, cost economies, and effect on overall food quality for
consumer acceptance require a combination of these processes to achieve the overall objective
of food preservation. Innovative non-thermal technologies that generated interest of the food
industry in recent times are ionizing radiation (gamma rays, beta rays, and pulsed X-rays),
application of pressure (high-pressure processing), application of electricity (pulsed electric
field: PEF), thermal radiation (ultraviolet [UV] light, pulsed light), and the magnetic fields
(oscillating magnetic fields). Commercial applications of these non-thermal technologies,
when applied individually, are only partially able to replace conventional thermal processes,
to meet the overall objective of food preservation. Any non-thermal processing technology
alone is not adequate to achieve commercial sterility, as resistant spores of some species are
found to survive after these treatments. This limitation can be overcome by combining two
or more non-thermal processing technologies and/or with traditional preservation techniques
to exploit synergistic interactions so as to achieve a hurdle effect. In recent times, hurdle
technology is being exploited as an integrated approach of basic food preservation. Hurdle
approach though is quite old in food processing, but with time, advances are being made and
its application is now concentrated with a multi-target approach by addition of new hurdles
(physical hurdles such as non-thermal technologies, chemical hurdles such as addition of
natural preservatives, etc.).
17.2 CONCEpt OF hUrDLE EFFECt
The hurdle concept is also known as the combination of preservation or barrier technol-
ogy or multifactorial preservation. It illustrates that preservation treatments can be combined
at lower individual intensities so as to enhance antimicrobial effects, while their impact on
sensory and nutritive properties of the food is minimal (Leistner, 1992). The hurdle concept
is well adapted in conventional or traditional preservation treatments, where low-intensity or
mild treatments were applied in synergy to achieve minimum degradation to nutritional and
sensory quality. Food is a complex matrix; therefore, each and every component needs to be
studied extensively to understand the mechanism of action of any technology applied. Also,
the successful application of hurdle technology requires knowledge of the mechanisms of each
hurdle applied (Leistner, 2000); this knowledge is lacking in the area of non-thermal process-
ing technologies, where it is essential to understand the synergistic mechanism on microbial
inactivation kinetics. Successful application of the hurdle concept in non-thermal food process-
ing technology requires intelligent selection of the mix of barriers (hurdles) and identification
of critical control parameters with respect to applied technology such as pressure, voltage,
dose, temperature, etc.
In food preservation, the word “hurdle” entails apparently a sequence of effects which forms
the basis of the successful hurdle technologies. Hurdle technology employs the use of different
hurdles that are deliberately combined in the preservation of conventional and novel foods
(Leistner, 1992). Initially hurdle technology was aimed to address the food safety in terms of
microbial quality, but with further developments in utilization of hurdle technology for food
preservation, significant weight was given to other parameters such as sensory and nutritional
quality in the optimization of food processes. By carefully selecting the suitable barriers in hurdle
combination, it is possible to address microbial stability and safety, sensory and nutritional quality
simultaneously, keeping economic aspects in view. When all the hurdles working in a particular
food are known, the microbial stability and safety of that food might be optimized by altering
the intensity or the nature of these hurdles (Leistner, 1999). Since about the last two decades, the
intelligent application of hurdle technology became more prevalent, because the principles of
major preservative factors for foods (e.g., temperature, pH, aw, Eh, competitive flora), and their
interactions, became better known.
Hurdles are differentiated and categorized into physical, physicochemical, microorganism-
derived, and other hurdles as described in Table 17.1. Therefore, substantial opportunities lie in
the smart use of a wider range of hurdles, both in combinations of non-thermal technologies and
in combination of non-thermal technologies with conventional preservation techniques. The use of
various combinations is limited by the preferred features in the final product as a result; a careful
selection of preservation factors (hurdles) is required due to the present demand of minimally
processed foods. Also, the excessive use or overloading of chemical preservatives or additives
is not preferred as it deviates from the main objective of minimally processed foods. The hurdle
approach is focused on development of minimally processed foods in industrialized nations, while
in developing nations the primary target is to keep food products stable as well as organoleptically
sound for longer periods without refrigeration (Leistner, 2000).
Further developments in hurdle technology led to the emergence of a novel concept of multi-
target food preservation, in which the influence of food preservation methods on the physiology and
behaviour of microorganisms in foods, i.e., their homeostasis, metabolic exhaustion, stress reactions,
are considered. Multi-targeted approach is the essence of hurdle technology, where, instead of
additive effect a synergistic effect could be achieved if the hurdles in a food, strike different targets
within the microbial cells (e.g., cell membrane, DNA, enzyme systems, pH buffering system, aw,
Eh, etc.), at the same time, and thus, disturb the homeostasis of microorganisms. Consequently,
making the restoration of homeostasis as well as the activation of “stress shock proteins” more
difficult. Therefore, employing simultaneously different hurdles in the preservation of a particular
table 17.1 hurdles Used for traditional and Modern Food preservation
physical hurdles
Conventional hurdles: High-temperature process (pasteurization, sterilization, blanching), low temperature
(refrigeration and freezing)
non-conventional hurdles: microwave, radio frequency, infrared and ohmic heating
Packaging hurdles: Aseptic packaging, vacuum packaging, modified atmospheres, controlled atmospheres,
active packaging, edible coatings
non-thermal hurdles: uV radiation, ionic radiation, electron beam radiation, ultrahigh pressures,
ultrasonication, PeFs, cold plasma, pulsed magnetic fields, electromagnetic energy.
physicochemical hurdles
Conventional hurdles: low pH, low redox potential, low water activity, smoking
natural antimicrobial systems: lactoperoxidase, lysozymes, lactoferrin, avidin
natural preservatives: spices and herbs, eos
traditional preservatives: salt, sugar
Artificial preservatives: organic acids, nitrite/ nitrate, sodium, or potassium sulphite, phosphates, carbon
dioxide, oxygen, ozone, ethanol, phenols, etc.
Microbiological hurdles
bacteriocins, competitive flora, protective cultures, antibiotics
Other hurdles
sanitizers, surfactants, oxidation products, chelating agents, etc.
food should lead to optimal stability (Leistner, 1995). The multi-target preservation of foods is
a potential research area, because if mild hurdles with different (multi) targets are selected, a
minimal but most effective preservation of foods could be accomplished. The application of multi-
target approach of hurdle technology in non-thermal processing technologies has created a new
line of interest to produce minimally processed, fresh-like food products that are stable at ambient
temperature for considerable time.
The use of non-thermal processes in combination with other preservation technologies presents a
number of potential benefits to food preservation. Combining non-thermal methods with other food
preservation techniques can (1) improve the lethal effects of non-thermal processing, (2) reduce the
severity of individual non-thermal treatment needed to obtain a given level of microbial inactivation,
and/or (3) prevent the proliferation of survivors following treatment. Considerable research
investigations have been carried out to explore the additive or synergistic outcomes of non-thermal
processing technologies with thermal processing, packaging technologies, antimicrobial agents, and
chemical preservatives. However, the research on combining two or more non-thermal processing
technologies in order to achieve synergistic effects is in its infancy stage. To expand the use of non-
thermal processes in the food industry, combinations of these technologies are being studied. Some
of the investigations where hurdle concept was used in combining non-thermal technologies and
traditional preservative factors are combination of thermal treatment with PEF (thermo-electrical
treatment), pressure (mano-thermal treatment), ultrasound (thermo-ultrasonication), irradiation
(thermo-radiation), and other hurdles (Kim and Thayer, 1996; Kalchayanand et al., 1998a; 1998b;
Raso et al., 1998b, 1998c; Marquenie et al., 2003; Gayán et al., 2014; ). Synergistic bactericidal
effects have also been observed between heat and ultrasound (Ordonez et al., 1987), heat and
radiation (Schaffner et al., 1989), high hydrostatic pressure (HHP) and nisin (Ponce et al., 1998),
and PEF and nisin (Calderon-Miranda et al., 1999). The combination processes involving non-
thermal technologies and conventional thermal processes, which have been patented to achieve
commercial sterility, are mano-thermo-sonication (MTS) and pressure-assisted thermal sterilization
(PATS). The combined effect of non-thermal technologies and conventional preservation techniques
are summarized in Table 17.2.
Since non-thermal technologies alone may not necessarily achieve commercial sterility,
continuous advances are being made to make these processes effective from commercial safety view.
table 17.2 Combined Effect of Non-thermal technologies

and Conventional preservation techniques
Conventional hurdles
Non-thermal technology heat ph antimicrobials
High hydrostatic pressure +++ ± ++/+++
Pulsed electric field +++ ± +++
ultrasonication +++ --- ++/+++
Continuous ultraviolet +++ --- ++
Pulsed light technology +++ --- ++/+++
ozonation +++ --- ++
irradiation --- ---
electron beam technology --- --- ++/+++,*
+ no Change in effect
++ Additive effect
+++ synergistic effect
Ântagonistic effect
±Mixed results
*Adverse effects on sensory Properties
These advances involve hurdle-technology approach based upon meticulous understanding of the
physics and chemistry involved in their mode of operation so that these processes can be exactly
defined and controlled. At present, the research activities in the area of hurdle technology focus
on discovering novel physical, non-thermal, and natural additives as hurdles. Hurdle technology
is an open thought that can easily be tailored to satisfy the desired objectives. Latest approaches
to hurdle technology in food sector include sustainable processing, predictive modelling, and food
packaging (Leistner and Gould, 2002).
For the advanced design of hurdle-technology foods, a 10-step procedure comprising hurdle
technology, predictive microbiology, and HACCP or GMP has been suggested by Leistner and
Gould (2002) as described in Table 17.3. Earlier, the type and intensity of hurdle was selected on the
ability to inhibit the microbial growth, but current researches emphasize selection of hurdles on the
principal of multi-target preservation.
Some of the research work assessed the potential for combining non-thermal technologies in
hurdle preservation approach. These combinations include UV and PEF (Noci et al., 2008), UV and
sonication (Char et al., 2010; Caminiti et al., 2011, 2012; Santhirasegaram et al., 2015a, 2015b), UV
and high-intensity light pulses (Caminiti et al., 2011), UV and radiofrequency electric field (Ukuku and
Geveke, 2010), pulsed light and ultrasound (Ferrario et al., 2015), high-intensity light pulses and PEF
(Caminiti et al., 2011), PEFs and ultrasonication (US) (Su et al., 1996; Jin et al., 1998), HHP and US
(Sala et al., 1995; Raso et al., 1998a; Pagan et al., 1999), HHP and modified atmospheres (Amanatidou
et al., 2000, Lopez-Caballero et al., 2000), HHP and PEF (Shimada and Shimahara, 1991; Pagan et al.,
1998; Shayanfar et al., 2014), irradiation and modified atmospheres (Lee et al., 1996; Hagenmaier and
Baker, 1998; Murano et al., 1998; Prakash et al., 2000), and irradiation and HHP (Sale et al., 1970;
Crawford et al., 1996). The combined effects of different non-thermal technologies are summarized
in Table 17.4.
table 17.3 Steps for Food Design Using an Integrated approach Comprising hurdle technology,
predictive Microbiology, and haCCp or GMp
1. For a modified or new food product the desired sensoric properties and shelf life are tentatively defined.
2. A feasible technology for the production of the food is outlined.
3. the food is manufactured, first at laboratory or pilot-plant scale, according to this technology, and the
resulting product is analyzed for pH, aw, preservatives, or other inhibitory factors. temperatures for
heating (if intended) and storage as well as the anticipated shelf life are defined.
4. For preliminary microbial stability testing of the food product, predictive microbiology is employed.
5. the product is then challenged with relevant food poisoning and spoilage microorganisms, using somewhat
higher inocula and storage temperatures than would be “normal” for the food.
6. if appropriate, the hurdles in the product are modified, taking multi-target preservation and sensory and
nutritional quality of the food (Le., total quality) into consideration.
7. the food is again challenged with relevant microorganisms, and if necessary the hurdles in the food are
modified again. Predictive microbiology for assessing the safety of the food is helpful at this stage too.
8. After the established hurdles of the modified or new foods are exactly defined, including tolerances, the
methods for monitoring the process are agreed on. Preferably physical or sensorial methods for
monitoring should be used.
9. the designed food should now be produced under industrial conditions, because the adequacy for a scale-up
of the proposed manufacturing process must be validated.
10. if, for an industrial process, the CCPs and their monitoring are established, the manufacturing process
might be controlled by HACCP. if HACCP seems inappropriate, guidelines for the application of
manufacturing control by quantitative GMP must be defined.
Adapted from Leistner, L. and G.W. Gould, Hurdle Technologies: Combination Treatments for Food Stability, Safety
and Quality. Kluwer Academic/Plenum Publishers, new york, p. 60, 2002.
table 17.4 Combined preservation Effect of Different Non-thermal technologies
Non-thermal hurdles
high hydrostatic pulsed electric Continuous pulsed light Supercritical
Non-thermal technology pressure field Ultrasonication Ultraviolet technology Ozonation Irradiation CO2 Microfiltration
High hydrostatic pressure ___ +++ +++ nL nL ++ +++ ++ +++
Pulsed electric field +++ ___ +++ +++ +++ ++/+++ nL +++ +++
ultrasonication +++ +++ ___ +++ ++/+++ +++ +++ +++
Continuous ultraviolet nL +++ +++ ___ nL +++ nL nL nL
ozonation ++ ++/+++ +++ +++ nL ___ nL nL nL
irradiation +++ +++ nL nL nL ___ nL nL
+ no change in effect
++ Additive effect
+++ synergistic effect
Ântagonistic effect
±Mixed results
*Adverse effects on sensory properties
nL no literature available
CoMbinAtion oF non-tHerMAL ProCesses AnD tHeir HurDLe eFFeCt
335
17.3 NON-thErMaL prOCESSING tEChNIQUES

IN COMBINatION WIth CONVENtIONaL hUrDLES
All kinds of hurdles, physical, chemical, physicochemical, or microorganism-derived, and mis-

cellaneous hurdles govern the growth of microbial cells. Factors that most effectively influence the
survival and growth of microorganisms are classified by Mossel (1983). They include:
Intrinsic factors: Physical and chemical factors that are within a food, and with which contaminating
microorganisms are inextricably in contact.
Processing factors: Procedures that are deliberately applied to a food in order to achieve improved
preservation.
Extrinsic factors: Factors that influence microorganisms in foods, but which are applied from, or exist,
outside the food, and act during storage.
Implicit factors: Those factors that are related to the nature of the microorganisms that are present, and to
the interactions between them and with the environment with which they are in contact during growth.
Net effects: Considering the net effects of the non-thermal technologies in combination, it is envisaged
that many of the factors strongly influence the effects of each other; however, the overall effects
of combinations of factors may not be obviously predictable, but may be usually greater than the
perceived effects of the single factors.
Initially, elevated temperature (achieved by mild thermal treatment), low temperature (achieved by
refrigeration storage), reduced pH (achieved by fermentation or by the addition of acids), and reduced
aw (achieved by partial drying or addition of solutes) have been the key contributors to successful
applications of hurdle technology. Later, antimicrobial efficacies have been steadily improved in
newer applications by the use of O2 free packaging and CO2-enriched modified atmosphere (Leistner
and Gould, 2002). Among the recent developments the amalgamation of bacteriocins, plant-derived
antimicrobials with HHP, PEF, MAP techniques in the course of hurdle technology to develop
minimally processed foods with considerable high shelf-life, optimum levels of nutrients and superior
sensory characteristics are being employed. Latest trend is toward the use of measures that produce
food products that are less heavily preserved and perceived as being natural or fresh-like, along with
high nutritional and sensory quality. Consequently, combination effects of non-thermal processing
technologies are the focus of many of the recent developments in the area of food sterilization. Due
to varied microbial resistance towards non-thermal technologies, sometimes very intense treatments
are required for assuring food safety and stability. This in turn possibly induces adverse effects on
sensory, nutritional, and functional quality of food product. Consequently, achieving synergistic or
additive effect in microbial reduction by combining non-thermal technologies with other barriers,
are in vogue. These emerging non-thermal techniques and their interactive effects which are among
the advanced research focus in area of non-thermal processing has been reviewed and limitations in
industrial scaling up of these combinations are also discussed.
17.3.1 high hydrostatic pressure in Combination with

Conventional preservation technologies
HHP processing employs the application of static pressures in the range of 100–1000 MPa to solid
or packaged foods, at ambient or near-ambient temperatures, from a several-second pulse to over few
minutes, in a batch or semi-continuous processing systems (Mertens and Deplace, 1993; Morris et al.,
2007). The process parameters such as pressure, temperature, rate of pressure change, treatment time,
and mode of pressurization (single or multi cycles) are the deciding factors towards inactivation of
microorganisms (Cullen et al., 2012). The mechanism of microbial destruction by HHP is based on
causing damage to the cell membrane. The lethal effect of high pressure on microorganisms is thought
to be the result of multi-target action taking place concurrently, involving breakage of non-covalent
bonds, damage to the cell membrane and inactivation of key enzymes (involved in DNA replication
and transcription). As HHP only involves breakage of non-covalent bonds leaving covalent bonds
intact, as a result low-molecular-weight molecules remain unaffected by application of high pressure
and hence maintaining the sensory and nutritional qualities of the food material. Furthermore, high
pressure disrupts secondary and tertiary structures of macro-molecules (proteins and polysaccharides)
which ultimately alter the functional properties of these macromolecules. Microorganisms illustrate
varied degree of resistance towards HHP (Stewart et al., 2000). Prokaryotes are more baro-tolerant
than eukaryotes; yeast cells are most sensitive among all microbial cells. Owing to structural complex-
ity in cell membrane of gram-negative bacteria, they are likely to be more susceptible to environmen-
tal changes caused by pressure than gram-positive bacteria. Vegetative forms of microbial cells are
sensitive than spores. Mould spores show evidence of certain degree of resistance to high pressures,
while bacterial spores exhibit extreme degree of resistance to the pressures normally applied to foods
with existence of significant variation between different species and strains. The extreme resistance
of bacterial spores encourages the requirement of intense treatment which otherwise is likely to affect
sensory and functional properties of food. Alternatively, to avoid side effects of extreme treatment lev-
els, HHP can be coupled with other preservation technologies to achieve desired goal of food safety.
HHP is the most extensively investigated in last few years including hurdle approach with different
conventional and emerging technologies with varied extent of results. A combination of different barri-
ers (preservation treatments), would most likely achieve food safety goals by effectively utilizing HHP
from commercial viability aspect.
17.3.1.1 Combination of High Hydrostatic Pressure with Acidification (Low pH)
HHP display different mechanisms for inactivation of vegetative cells and spores and so as
its combination with other barriers exhibit diversity in behaviour. It is thought that the mode of
action of the combination of HHP with acidification in vegetative cells generate inhibitory effect
on membrane ATP-ase, which is essential for intra-cytoplasmic pH regulation of vegetative cells
(Hoover et al., 1989); while, in spores, acidic media might aggravate a loss of divalent cations from
the cortex or its protonization, resulting in cortex alteration and subsequent protoplast rehydration
which ultimately leads to loss of baro-tolerance.
Antimicrobial effect of HHP in combination with low pH have shown mixed results, where,
some studies have shown that the antimicrobial effect of HHP can be increased when used in
combination with low pH while others have depicted positive effect at neutral pH. A review reported
by Raso and Barbosa-Cánovas (2003) also states microbial inactivation of both vegetative cells and
bacterial spores with HHP is barely affected by the pH of the treatment medium.
Roberts and Hoover (1996) reported an appreciable microbial inactivation by an additional
1.5 log10 in a buffer system when the pH was lowered from 7.0 to 4.0, which may be attributed to the
increase in the effectiveness of pressurization in presence of reduced pH. In another study, it was
reported that pressurization at 404 MPa (25°C for 15 or 30 min) at pH 4.0–7.0 resulted in a 0.5-log
reduction of Bacillus subtilis spore suspensions with initial concentrations of 106 spores/ml while
in spores of Clostridium sporogenes PA 3679, a reduction of 2.5 log10 –3 log10 was achieved at pH
4.0 (Stewart et al., 2000). Sale et al. (1970) illustrated that spores were inactivated by pressure over
a wide pH range but a greater inactivation was observed near neutrality than at extreme pH values.
Balasubramaniam et al. (2001) achieved an additional reduction of 2.5 log of B. subtilis spores at
827 MPa and 75°C, when the pH of the treatment buffer was reduced from 7.0 to 3.0. Conversely,
some authors have reported that varying pH has modest effect on HHP inactivation of yeast and
bacterial spores (Patterson et al., 1995; Raso et al., 1998b).
Ogawa et al. (1990) concluded that the inactivation effect of pressure does not depend on the pH of
the Satsuma mandarin juice. When, the effect of pH on the pressure inactivation was tested in the juice
with the adjusted pH from pH 2.5 to 3.5 using different organic acids (citric, malic, tartaric, lactic, or
acetic acid). The result shows that pH does not influence the pressure-induced inactivation with selected
species of yeasts and moulds. On the other hand, Pandya et al. (1995) showed that pH has a marked
effect on destruction of Saccharomyces cerevisiae and Zygosaccharomyces bailii when exposed to
2,500 atmospheres at 25°C for 20 min. The results expressed a 7.0 log10 cycle reduction at pH 3.0 and
4.0 as compared to 4.0 log10 cycle at pH 5.0. Garcia-Graells et al. (1998) also indicated an increase in
the rate of inactivation at the reduced pH under pressurization, when pressure resistance at low pH were
compared for their inactivation rate during storage in different juices (apple, orange, or mango) at the
same pressure (300 MPa), or in the same juice at different pressures (300 and 400 MPa).
Timson and Short (1965) concluded that the pressure resistance of the organism decreases as the
pH is lowered or raised, a slightly greater inactivation of Bacillus subtilis was observed at pH 6 than
at pH 8. Linton (1999) established that pH has a significant effect on inactivation rates of Escherichia
coli O157H:7 in orange juice during storage. The orange juice was pressurized at 400 MPa for 1 min
at 10°C over the pH range of 3.4 to 5.0. It was observed that the use of high-pressure processing of
orange juice made E. coli O157:H7 susceptible to the high-acid conditions found in orange juice.
Pressurization in the presence of either citric or lactic acid increased the viability loss by an additional
1.2–3.9 log cycles at pH 4.5 for both acids at 345 MPa suggesting possible pressure pasteurization
applications to liquid foods that have low pH using this combination of treatments (Alpas et al., 2000).
17.3.1.2 High Hydrostatic Pressure in Combination with Heat
This combination of treatments in non-thermal processing of foods for its anti-microbial effects
has been widely studied by researchers. Application of HHP has been found to increase the destruc-
tion of vegetative cells of microorganisms at temperatures below or above ambient temperature
(30°C) (Mackey and Manas, 2008). At low temperatures (refrigeration or freezing), the antimicrobial
effect is associated with the susceptibility of proteins to denaturation and an irreversible decrease in
membrane fluidity, while at higher temperatures (above 30°C–35°C) the effect is due to phase transi-
tion of membrane lipids (Kalchayanand et al., 1998a; Mackey and Manas, 2008). It is also evident
from several research outcomes that HHP with simultaneous application of moderate heat is more
effective for inactivation than their additive effect; therefore, this effect is stated as synergistic. The
destructive effect of HHP and heat might associate with the cellular injury due to oxidative damage
in cells (Aertsen et al., 2005). Much less deviation was reported in pressure resistance between dif-
ferent microbial species or between different strains of the same species in application of combined
treatment of HHP with temperature (Alpas et al., 1999). Bacterial spores are more baro-tolerant and
some species are even unable to inactivate at pressures as high as 1200 MPa, therefore, spores could
not be inactivated by pressure alone (Gould and Sale, 1970; Mills et al., 1998). A two-stage process
of germination and inactivation of microbial spores is followed in HHP processing when combined
with heat. A mild pressure of even 10 MPa have been found to activate spore germination (Gould and
Sale, 1970), while lethality (instigation of germination and inactivation of bacterial spores) of HHP
at elevated temperatures is greatly enhanced (Raso et al., 1998a). As pressure-germinated spores lose
their resistance to physico-chemical agents, the germinated spores may be inactivated at significantly
low-pressure levels or with other mild treatments than in case of spore. Conversely, existence of a
comparatively large portion of super dormant spores that remain ungerminated after lengthened
pressurization, even in presence of mild heat, needs considerable attention (Sale et al., 1970).
Carlez et al. (1993) depicted efficient reduction (Citrobactor freundii, Pseudomonas fluorescens, and
Listeria innocua) in minced beef could be achieved by use of lower intensities of pressure (200 MPa in
case of gram-negative and 350–450 MPa in case of gram-positive) at refrigeration temperature of 4°C
or under moderate heating at 35°C–50°C. Combination treatments of heat and pressure when applied
simultaneously (e.g., 400 MPa at 60°C for 30 min) or sequential (e.g., 80°C for 10 min followed by
400 MPa for 30 min) mode, proved more effective in inactivating spores of Clostridium sporogenes,
which were resistant to inactivation at 600 MPa for 30 min at 20°C (Mills et al., 1998).
Vegetative forms of yeast and moulds are susceptible to high pressures at ambient temperature;
therefore, its combination with heat is hardly studied. Whereas, mould ascospores have expressed
certain degree of resistant to high pressures and needs mild heat in combination for inactivation.
Pressure sensitizes ascospores to subsequent heat treatment. B. nivea ascospores require pressures
above 600 MPa and temperatures above 60°C for inactivation. Treatment of 700 MPa at 60°C
sensitizes B. nivea ascospores to subsequent thermal treatment (Butz et al., 1995). A combination of
high pressure and moderate temperature has proved very successful in microbial inactivation. The
cell death and injury of the four pathogens (Staphylococcus aureus 582, Listeria monocytogenes
scott A, Escherichia coli 0157:H7 932, and Salmonella typhimurium ATCC 14028) as affected by
pressurization temperature were studied by exposing the cells at 207 MPa for 25°C–45°C and it was
concluded that proportionately much higher cell death was achieved above 35°C (Kalchayanand
et al., 1998a). Hayakawa et al. (1994) observed an additional 2 log cycles inactivation of
B. stearothermophilus with combination of an oscillatory pressurization (6 cycles of 5 min/
oscillation, 400 MPa at 70°C) and heat as compared to HHP (600 MPa, at 70°C) and heat alone. Six
times oscillatory pressurization under 600 MPa at 70°C achieved complete sterilization.
Effect of HHP on enzymes has shown varied responses which may be attributed to the struc-
tural differences of individual enzymes. In majority of studies, combinations of pressure with mild
heat found to increase the level of enzyme inactivation, but in some cases an increase in enzyme
activity has been reported. HHP treatment induces conformational changes owing to protein dena-
turation that can change the functionality of the enzyme (e.g., increase or loss of biological activity,
change in substrate specificity). In addition to structural changes, enzyme activation by HHP might
be linked with pressure-induced membrane damage and the resulting interface of enzyme and
substrate due to decompartmentalization while, in intact tissues, enzymes and substrate are often
separated in screened-off area, which is easy to act upon by application of low pressure (Hendrickx
et al., 1998).
A study on the effect of HHP treatment (50–500 MPa) in combination with moderate heat
treatment (20°C–60°C) on peroxidase (POD), polyphenol oxidase (PPO), and pectinmethylesterase
(PME) activities of tomato puree revealed a combination of 150 MPa/30°C produced 32.5% reduction
of PME activity, while some activation was observed at 335–500 MPa and different temperatures.
A reduction of 25% POD activity was obtained at 350 MPa/20°C, whereas a combination of higher
pressures and mild temperatures (30°C–60°C) produced an enhancement of this activity. PPO activity
was not affected by UHP/mild-temperature treatments and only a combination of 200 MPa/20°C
produced a significant loss (10%) in PPO activity (Hernandez and Cano, 1998). In pressure-treated
potato cubes (400 MPa, 15 min) limited inactivation of PPO was observed at 5°C, whereas a
significant inactivation was achieved at increased temperature (Eshtiaghi and Knorr, 1993).
Further advancements envisaged PATS as an alternative method for sterilization of food products.
PATS employs simultaneous application of elevated pressures (up to 900 MPa) and sub-retorting
temperatures (90°C–120°C) to a preheated food (75°C–95°C) (Balasubramanian and Balasubramaniam,
2003). In PATS, instantaneous increase in temperature of the tested food product is attributed to
heat of compression due to rapid pressurization and subsequent cooling (up to initial temperature)
upon depressurization. The severity of thermal effects as come across during conventional thermal
processing was reduced by compression heating. Bacillus stearothermophilus inoculated egg patties
were preheated and then PATS treated at 105°C for varying degree of pressures and holding times.
Exposure of PATS at 700 MPa and 105°C resulted in 4 log reduction of B. stearothermophilus spores
in 5 min in comparison to 1.5 log reduction in 15 min thermal treatment at 121°C. Spore inactivation
by PATS progressed rapidly up to 100 s pressure-holding time, but greater holding times showed
comparatively limited effect (Rajan et al., 2006). The effect of PATS on microbial counts and on activity
of polyphenol oxidase enzyme was assessed and the results revealed that in PATS-treated purées, the
inactivation rate increased proportionally with pressure and temperature, at 80°C and 900 MPa the
reduction in microbial load was under detection limit along with reduction of viable spores of Bacillus
by at least in 2 log units. Almost all of the applied treatments reduced noticeably the PPO activity and
the highest reduction in activity was encountered at 600 MPa/70°C (García-Parra et al., 2016).
It is well established that a prolonged exposure of food product to intense heat or intense
pressures produces undesirable changes. Application of intense heat treatment destroys the heat
labile nutrients and affects the sensory quality while high pressures leads to protein denaturation
and textural modifications in food products. Therefore, combining moderate temperature and
pressure could be an effective combination to cope up with these drawbacks. Also, using less energy
intensive processes will improve the cost economies making the treatment commercially viable.
17.3.1.3 High Hydrostatic Pressure in Combination with Antimicrobials
Consumer’s demand for natural products is driving the current trend towards the use of natural
systems such as microbial antagonism (antimicrobials) in food preservation. The natural antimicro-
bials possess generous potential when used in combination with other food preservation techniques
(Gould, 1996). The natural antimicrobials are categorized as: plant derived (herb and spices extracts),
animal derived (lysozyme, lactoperoxidase system (LPS), lactoferrin, lactoferricin), products of
microorganisms (bacteriocins such as nisin, pediocin other bacteriocins and culture products anti-
mycotics, natamycin/pimaricin) (Leistner and Gould, 2002). In wide-ranging, bacteriocins cannot be
used as a sole means of preservation to prevent the augmentation of pathogens but can be presented as
a supplementary barrier to reduce the odds of food borne disease. The inactivation of gram-negative
bacteria is a matter of concern while using antimicrobials such as bacteriocins. The resistance is
due to protective outer membrane which provides an efficient barrier against hydrophobic solutes
and macromolecules, such as bacteriocins (Halander et al., 1997). Therefore, hurdle approach is an
important way to utilize antimicrobials in combination with HHP to obtain a synergistic effect for
inactivation, as HHP disrupts the membrane integrity at lower pressures applied. The combination
of antimicrobials with HHP treatment has been proposed to achieve a high degree of inactivation of
gram-negative bacteria. Also, as applications of bacteriocins in various food matrices are still being
discovered, consequently in near outlook addition of newer bacteriocins with more value and potency
to existing class is expected.
It was evident from the findings that, high pressure (>180 MPa) exposure showed an increased
lethality in the presence of lysozyme, nisin, and ethylene diamine tetra acetic acid (EDTA). It
was found that sub-lethal injury in the surviving population was lower in the existence of nisin
and lysozyme, but higher in the presence of EDTA. An additive effect in lethality was observed
in combinations of EDTA with nisin or lysozyme. However, the addition of lysozyme, nisin and/
or EDTA to pressure-treated cell suspensions instantaneously after pressure treatment did not
result in effective microbial reduction Hauben et al. (1996). Masschalck et al. (2000) reported that
upon application of pressure, both lysozyme and nisin exhibited enhanced lethality but the level
varied between strains. Nisin improved pressure inactivation of all the mutants by at least 2 logs
for pressures 300 MPa. Similarly, combined pressure lysozyme treatment resulted in an increased
lethality of approximately 2 logs for pressures 300 MPa except in case of mutant LMM1010. Garcia-
Graells et al. (1999) depicted an additional reduction of 3 logs in skim milk at 550 MPa when
coupled with lysozyme (400 μg/mL) and nisin (400 IU/mL) before pressure treatment. The increase
in lethality was ascribed by enhanced sensitivity of the cells to high pressure as well as to the action
of lysozyme and nisin. However, the effect was dependent on strain and composition of milk.
Additional viability loss in between 1.3 and 5.1 log cycles in four pathogens (Staphylococcus
aureus 582, Listeria monocytogenes scott A, Escherichia coli 0157:H7 932 and Salmonella
typhimurium ATCC 14028) was acquired, when a mixture of pediosin AcH and nisin A coupled
with pressurization for 10 min was applied (Kalchayanand et al., 1998b). Combination of HHP
with nisin established a reduction of almost 5 log10 units in E. coli counts and more than 6 log units
for L. innocua at 450 MPa and 5 mg/L of nisin in liquid whole egg (Ponce et al., 1998).
In an investigation, a natural antimicrobial system, the LPS, consisting of lactoperoxidase, thiocyanate

(SCN−) and hydrogen peroxide (H2O2) present in milk was evaluated for its effectiveness in combination
with pressure. The use of LPS has been established as an effective method to extend the shelf-life of raw
milk. The bactericidal action of H2O2 is attributed to the generation of antibacterial products such as
hypothiocyanite (OSCN−) anion and hypothiocyanous (HOSCN) acid due to the oxidation of SCN− by
H2O2. These short-lived reaction products are known to cause oxidation of sulfhydryl groups of bacterial
cell membrane’s proteins and inhibit several important enzymes in cellular metabolism, interferes with
the transport of nutrients, the DNA and RNA synthesis and the respiratory chain (Naidu, 2000). The
combined effect of HHP (450 MPa, for 10 min) and LPS on the inactivation of Listeria monocytogenes
H66a in cold-smoked salmon during 35 days at 5°C depicted synergistic antimicrobial effect, preventing
the pathogen recovery, suggesting potential combination against L. monocytogenes, increasing the safety
and the shelf-life during refrigerated storage (Montiel et al., 2012).
In line of use of natural antimicrobials, synergism in inactivation rate was also reported to be
achieved by combined processes of HHP and essential oils (EOs) or their chemical constituents
(CCs) on Escherichia coli O157:H7 and Listeria monocytogenes EGD-e. The tested combinations
expressed varied responses towards inactivation rate, the most effective being (+)-limonene,
carvacrol, C. reticulata L. EO, T. algeriensis L. EO and C. sinensis L. EO which resulted in about
4–5 log10 cycles of the initial cell populations in combination with HHP indicating an outstanding
synergistic effect. The encouraging outcome suggests possible use of this combination in liquid
food such as fruit juices (Espina et al., 2013).
Combining antimicrobials with HHP enhances the inactivation effect of the HHP and broadens
the spectrum of action of some natural antimicrobials with advantages in product safety. However, the
inhibiting effect of the antimicrobials on the growth of the survived microorganisms following the
treatment has not been investigated and needs to be taken to next level of research investigations.
17.3.2 Ultrasonication in Combination with Conventional

preservation technologies
US employs the application of sound waves (vibrational energy with continuous frequencies)
with frequencies exceeding the hearing range of human ear (~20 kHz). Ultrasound application in
food industry is divided into two categories on the basis of frequency range applied. Low energy
(<1 W·cm2), high-frequency (>100 kHz) ultrasound are utilized for non-destructive food testing and
monitoring during processing and storage, while, high energy (intensity >1 W·cm2), low frequency
(between 20 and 500 kHz) are disruptive in nature and stimulate effects on the physical, mechani-
cal, or chemical/biochemical properties of foods (Awad et al., 2012). The application of high-power
ultrasound (HPU) or power ultrasound is of more importance in extending shelf-life by microbial
inactivation. The mechanism of microbial destruction is associated with thinning of cell membranes,
localized heating, and production of free radicals due to cavitation phenomenon (Butz and Tauscher,
2002). The important parameters affecting the effectiveness of ultrasonic waves towards microbial
inactivation are amplitude of the ultrasonic waves, exposure time, volume of food being processed,
the composition of the food material and the treatment temperature (USDA, 2000). According to
several research findings, the sensitivity of different forms of microbial cells towards ultrasonic
treatment varies in following order: bigger cell > smaller cell, rod-shaped bacteria > coccal forms,
gram-negative > gram-positive, anaerobic species > aerobic, young cells > old cells, vegetative cells
> sporulated microorganisms (Ahmed and Russell, 1975).
In a research investigation, a change in target of ultrasonic damages has been observed with the
change in intensity (low and high). At low acoustic intensity (10 W/cm2) quicker release of cell wall
polysaccharides than the intracellular proteins was observed, whereas; at higher acoustic intensities
(24 and 39 W/cm2) a reverse trend was observed. The disruption of yeast cells at low acoustic
intensities was associated with the initiation of the breakdown of the cell wall prior to damage of the
cell membrane (Wu et al., 2015). These findings may play an important role in efficient application
of US as a preservation technique. US have proven to be a potential preservation technique for
use in the beverage industry (Knorr et al., 2011); however, inactivation efficiency of ultrasonic
waves with definite frequency or power depends on types of microorganisms. Bacterial spores
are resistant to ultrasound treatment alone; therefore, like other non-thermal technologies a hurdle
approach is required to achieve desired objectives. The advantages of ultrasounds over conventional
pasteurization comprise of better economy, reduced flavour losses with high degree of homogeneity.
Several research findings have demonstrated the additive and synergistic effect of ultrasound
in conjunction with heat and/or pressure. The three combinations of combined effect vastly
studied are thermosonication (ultrasound + heat), manosonication (ultrasound + pressure), and
manothermosonication (ultrasound + pressure heat).
17.3.2.1 Ultrasonication in Combination with Heat (Thermo-sonication)
The combination of ultrasound and heat treatment is known as thermo-sonication (TS). Results of
numerous studies showed that ultrasound and temperature had synergistic effects on inactivation of
microorganisms. It was established that the microbiological cells became more sensitive to heat after
being exposed to ultrasonic waves, as elevated temperature weakens the cell wall, depolymerize macro-
molecules and thermally coagulates the intracellular proteins, however; this effect was observed only up
to a certain temperature limit, above which synergistic effect was found to diminish. This may possibly
be due to the cushioning effect of vapor in cavitation bubbles. When liquid media is exposed to ultrasonic
waves in conjunction with heat, temperature affects vapor pressure, surface tension, and viscosity of the
medium. With increasing temperature in the liquid medium, the equilibrium vapor pressure of the sys-
tem and the formation of cavitation bubbles increases. The presence of additional vapours in cavitation
bubbles creates cushioning effect preventing the implosion of the bubbles during cavitation and affecting
the microbial inactivation. In contrary to this effect, increased temperature decreases the viscosity of the
liquid medium, which leads to easier bubble formation. Consequently, there is an optimum temperature
for synergism between ultrasound and heat in liquid media (Sala et al., 1995). Also, an assured success
of TS treatment could only be achieved if variables such as viscosity, suspended solids content, pH in
the medium, acoustic energy density (frequency and amplitude) temperature, and time of the treatment
are well addressed (Guerrero et al., 2001). Several researchers have reported that ultrasound at atmo-
spheric pressure and controlled temperatures is effective against pathogenic and spoilage microorgan-
isms. However, some investigations suggested that ultrasound at a controlled temperature (25°C) may not
be effective for inactivation of some types of microorganisms and enzymes and recommended a better
effect when combined with moderate heat. A classification is also given to TS according to the applica-
tion of temperature used into sub-lethal (<45°C) and lethal (>45°C) treatment (Tiwari et al., 2009).
According to Ciccolini et al. (1997), ultrasonic waves alone could not destroy the S. cerevisiae
whereas increased inactivation was observed in combination with heat due to increased sensitivity
to temperature.
When raw whole milk was treated with ultrasound (0, 40, 72, 108, and 120 μm) in combination
with temperature (63°C) to inactivate Listeria innocua and mesophilic bacteria, a 5-log reduction
was achieved after 10 min as compared to 0.69 log after thermal pasteurization for same duration
(Bermúdez-Aguirre et al., 2009). The simultaneous application of ultrasonic waves (20 kHz, 150 W)
and heat treatment on the survival of a strain of Staphylococcus aureus in 0.05 M-phosphate buffer
(pH 6.8) and whole milk showed a 63% decline in decimal reduction times when the test organism
was suspended in buffer and 43% decline in decimal reduction times when suspended in milk as
compared to thermal treatment alone. The authors concluded that the effect was much greater than
the additive effect of the two agents when considered independently (Ordonez et al., 1987).
In general, studies show that TS is effective to attain vegetative microbial cells inactivation beyond
5 log cycles (Anaya-Esparza et al., 2017). Microbial spores are resistant to ultrasound treatment
alone, therefore, its combination with heat was studied and positive results were obtained. The TS
effect (20 KHz, 150 W and 70°C–95°C) on the survival of two strains of Bacillus subtilis in three
suspending media (distilled water, glycerol, and milk) was studied. It was reported that simultaneously
application of both the treatments decreased decimal reduction times by 79% (B. subtilis var. niger-40)
and 40% (B. subtilis ATCC 6051) in milk as compared to heat alone, whereas, no decrease of the heat
resistance was observed when ultrasonic was used as pre-treatment before heat treatment. Also, a
significant fall in the TS effect was accounted as the temperature of the treatment approached to the
boiling point of the water which is explained by cushioning effect (Garcia et al., 1989). The most heat
resistant Clostridium perfringens spores which can survive thermal treatment at 100°C for 1 h or more
(Labbé et al., 2014) showed reduced thermal resistance by half in beef slurry when heat shock treatment
at 80°C for 10 min was given followed by 1 min ultrasound treatment (Evelyn and Silva, 2015). Milani
and Silva (2017) showed promising outcomes of ultrasound-assisted thermal pasteurization of beer,
aiming to achieve the inactivation of Saccharomyces cerevisiae ascospores, the most resistant form of
the yeast. The batch operation mode showed effective inactivation as compared to continuous mode.
TS at 50°C for 1.9 min and at 55°C for 26 s were found adequate for pasteurization, in contrast to
55°C for 38 min in thermal processing. The researchers concluded with an effective combination of
treatments: TS at 50°C for 3.0, 1.9, and 4.5 min for the minimum requirement of pasteurization of beer
with 0.0, 4.8, and 7.0% alcohol/volume, respectively.
The effect of ultrasonic waves on enzymes depends on their molecular structure, sonicating
medium, and nature of the dissolved gas in addition to factors related to the ultrasonic processing.
Sometimes a loss of enzymatic function was observed due to change in catalytic functions while in
other findings no such effects have been reported (El’Piner, 1964). According to Rojas et al. (2016),
inactivation of POD in green coconut water showed inactivation of approximately 27% after 30 min of
processing, when ultrasound was used alone. Furthermore, US technology showed a positive impact
on consequent heat processing and a lower thermal resistance was observed. The change in thermal
resistance might be associated with the changes caused in the enzyme structure making the enzyme
population more homogeneous which in turn leads to more uniform distribution of resistance, allow-
ing lower processing time and/or temperature. TS treatment aiming to inactivate tomato PME showed
a strong synergistic effect with an increased inactivation by 39–374-fold at 61°C and 36–84 fold at
72°C, depending upon cavitation intensity. The increase in the inactivation was more pronounced at
low temperatures. Also, D-value was reported to reduce to 0.3 min in case of TS at 72°C as compared
to 25.3 min in thermal processing alone at same temperature (Raviyan et al., 2005).
17.3.2.2 Ultrasonication in Combination with Antimicrobials
The combination of US and antimicrobials expressed progressive results as a hurdle approach

towards achieving food safety and quality concurrently. Combination of ultrasound with natural
antimicrobials (vanillin and pomegranate extract) to obtain shelf-stable strawberry juice was inves-
tigated in order to find the optimum conditions for all three preservation hurdles. The optimal
conditions to simultaneously minimize native microflora, maximize nutritional parameters, and
minimize the impact on sensory quality resulted in 7.5 min of ultrasound treatment, pomegranate
extract concentration of 360 mg/mL and vanillin concentration of 0.925 mg/mL. This combination
of preservation hurdles showed promising results in extending the shelf-life of the product while
maintaining the sensory and nutritional quality (Tomadoni et al., 2016). Decontamination efficiency
of ultrasound in combination with EOs (oregano and thyme) in Salmonella inoculated lettuce leaves
was investigated. The experimental findings have shown less than 1 log CFU/sample inactivation in
total bacterial count when both the treatments applied individually. However, a synergetic effect on
inactivation was observed when a critical concentration (0.018% or higher) of EOs was simultane-
ously applied with ultrasound (Millan-Sango et al., 2016). Carvacrol or thymol at critical concentra-
tion is known to sensitize the bacteria by saturating the site of action of membrane which ultimately
subside the integrity of the bacterial cytoplasmic membrane and, hence, the leakage of vital intra-
cellular constituents (Juven et al., 1994).
Incubation of yeast cells with chitosan (1000 ppm) prior to sonication enhanced the inactivation of
Saccharomyces cerevisiae by ultrasound. Incubation with chitosan for 30 min resulted in approximately
1-log cycle reduction of the yeast, which leads to a final reduction of more than three log cycles
following 30 min of the ultrasonic treatment. Ninety minutes of incubation with chitosan resulted in
significant change in yeast reduction which might attributed to chitosan hydrolysis by enzymes and/
or development of protective envelop by yeast during the prolonged incubation (Guerrero et al., 2005).
The investigation reviewed the synergistic effects of ultrasound (37 kHz, for 5–100 min) and
sodium hypochlorite (NaOCl) (50–200 ppm) on inactivation rate of Cronobacter sakazakii KCTC
2949 in head lettuce. Ultrasound alone did not show progressive results with reduction of only 0.01–
0.58 log; whereas, NaOCl showed significant reduction of 0.58–2.77 log. Moreover, the combination
of ultrasound (100 min) and NaOCl (200 ppm) resulted in an additional 1.67-log reduction (Park et al.,
2016). The effectiveness of commercial sanitizers (sodium dichloroisocyanurate, hydrogen peroxide,
chlorine dioxide, peracetic acid) in conjugation with ultrasound for decontamination step of minimally
processed cherry tomatoes was investigated. For this purpose, the reduction of natural microbial load and
inoculated Salmonella adhered to the surface of the tomatoes were evaluated. The combined treatment
of ultrasound and 40 mg/L peracetic acid resulted in the highest reduction of the natural microbial load
and a reduction of adherent Salmonella typhimurium ATCC 14028 by 3.9 log10 cfu/g. These results
indicate the possibilities of using ultrasound as assisted approach towards decontamination of cherry
tomatoes to obtain minimally processed products (São José and Vanetti, 2012).
Effect of combining various hurdles in a model system on survival of Aspergillus flavus was stud-
ied by Coronel et al. (2011). Combining TS, natural antimicrobial (500 ppm vanillin) and reduced pH
(3.0) established efficacy in mould inactivation as short treatment times were required (2.5–3 min).
Use of combined ultrasound, natural antimicrobial and stress factors as pH allows using lower tem-
peratures and shorter treatment times for microbial inactivation in order to improve food quality.
17.3.3 Continuous Ultraviolet processing with possible hurdles
The non-thermal processing engaging the use of UV light is well recognized in food industry
for water treatment, surface decontamination and sterilization of food packaging material but its
application for treating liquid foods containing solutes or material in suspension is still limited due
to varied optical and physical properties and diverse chemical compositions of food products that
influence UV light transmittance, dose delivery, momentum transfer, and consequently microbial
inactivation (Koutchma, 2009). The inactivation mechanism of UV involves direct effect on nucleic
acid that absorbs UV light in the range of 200–310 nm resulting in creation of pyrimidine dimers
which prevent replication of microorganisms and hence making them inactive to cause any undesir-
able effect. UV disinfection system employs low or medium pressure mercury lamps for generation
of continuous UV radiation between 200 and 300 nm. Continuous UV radiation has limitations of
poor penetration depth and low emission power which to a certain extent is taken care of by advanced
version of UV processing known as pulsed light technology (PLT). However, as a non-thermal pro-
cessing technology, UV irradiation has an optimistic consumer response and is of significance to the
food industry as a low-cost alternative to thermal processing. The excessive use of UV radiation to
achieve desired inactivation in food products can lead to rancidity due to peroxidation of fats, furan
formation in fruit juices, and “brown spot” in certain types of fresh produce (Sharma 2014); there-
fore, minimization of these undesirable effects could be best achieved in combination with other
preservation technologies. UV radiation has been shown to have a synergistic disinfective effect
when used as a barrier in hurdle concept with other conventional and non-thermal processing tech-
nologies. The effect of UV radiation in combination with various conventional (heat and chemical
disinfectants) and non-thermal (sonication, PEF) preservation barriers is summarized in Table 17.5.
table 17.5 Combined treatments to Inactivate Microorganisms by Means of Continuous Ultraviolet and Other Conventional and Non-thermal technologies
hurdles process Conditions target Microorganism Log reductions Food Matrix references
uV-C+heat 27.1 mJ/L for 3.6 min+52.5°C enterotoxigenic 5.0 Fruit juices and Gayán et al. (2014)
or Staphylococcus aureus vegetable and
13.6 mJ/L for 1.8 min+55.0°C chicken broths
2.9 J/mL for 2.7 min+57.5°C Saccharomyces 5.0 Clarified apple juice Gouma et al. (2015)
cerevisiae
23.72 J/mL for 3.6 min+55°C Escherichia coli >5.0 orange juice Gayán et al. (2012)
uV-C+sodium benzoate 1.4 L/min Aspergillus flavus or up to 3.5 Peach nectar Flores-Cervantes et al.
Aspergillus niger (Did not achieve (2013)
5 log10 reduction)
uV-C+dimethyl dicarbonate 10.76 mJ/cm2 per cycle+250 total plate count and 2.61 Pineapple juice shamsudin et al.
ppm yeast and mould count 4.87 (2014),
(Did not achieve
5 log10 reduction)
uV-C+citric acid+dimethyl uV-C+1.5%+15 µL/100 Ml total plate count and 4.12 red pitaya juice Halim et al. (2012),
dicarbonate yeast and mould count 4.14
(Did not achieve
5 log10 reduction)
uV-C+aqueous Clo2+fumaric 1.9 kJ/m2 uV-C+100 ppm total aerobic bacteria 3.9 buckwheat sprout Chun and song (2014)
acid aqueous Clo2+0.31% fumaric yeast and mould 1.8
acid Coliform 2.4
uV-C+manothermosonication 5.3 J/cm2 for 30 s+20 kHz, 23 Escherichia coli and Pichia >6.0 Apple and cranberry Palgan et al. (2011),
μm with 400 kPa bars over fermentans juice blend
atmospheric pressure
uV-C+ultrasonication 0–18.7 kJ/m2 + 20 kHz, 95 µm E. coli strains 3.4 orange and/or apple Char et al. (2010)
simultaneous application for juices
20 min.
uV-C+ultrasonication 3.525 J/m2+40 kHz at 25°C for Coliform count not detectable Chokanan Mango santhirasegaram et al.
15 and 30 min) Juice (2015b)
Aerobic plate count not detectable
CoMbinAtion oF non-tHerMAL ProCesses AnD tHeir HurDLe eFFeCt
yeast and mould count 1.0

uV-C+PeF 5.3 J/cm2+34 kV/cm for 93 µs Escherichia coli and Pichia >6.0 Apple and cranberry Palgan et al. (2011)
fermentans juice blend
30 w, 30 min+100 μs, 40 kV/cm S. aureus 9.5 Apple juice Walkling-ribeiro et al.
(2008)
30 w, 30 min +100 μs, 40 kV/cm natural flora 7.1 Apple juice noci et al. (2008)
uV-C+ozone 0.9 g ozone/h at a flow rate of L. monocytogenes >9.0 Fresh brine Kumar et al. (2016)
2.4 L/min for 10 min +350–
400 mW/cm² uV for 10 min
60 min ozonation+10 minutes uV >5.0 spent brine
345
17.3.4 pulsed Light processing in Combination with

Conventional preservation hurdles
Pulsed light technique is known by several names in different research findings which include
pulsed UV light, pulsed white light, and high-intensity pulsed light. High-voltage and high-current
electric pulses as used in PEF processing are converted into short-duration and high-power pulses
of radiation included in the spectra of UV (180–400 nm), visible (400–700 nm), and infrared light
(700–1100 nm) by inert-gas flash lamps (Palmieri and Cacace, 2005). Although wide variations exist,
approximately 54%, 26%, and 20% of the emitted energy correspond to UV, visible, and Near-Infrared
light (NIR light), respectively. The UV portion of the electromagnetic spectrum includes long-wave
UV-A (320–400 nm), medium-wave UV-B (280–320 nm), and short-wave UV-C (200–280 nm), which
represent 25%, 8%, and 12% of the total emitted light, respectively (Condon et al., 2014). Pulsed light
processing employs application of short duration pulses of an intense broad spectrum which is rich in
UV-C light. UV-C portion of the electromagnetic spectrum corresponds to the wavelength between
200 and 280 nm. The duration of emitted light flash varied from millionths or thousandths of a second.
The most accepted mechanism for killing of microorganisms by PLT involves co-existence of
photochemical (lethal effects of light pulses) and photo-thermal (due to rapid temperature increase)
effect. The photochemical effect is attributed by the action of UV light directly on the DNA of
microbial cells leading to the formation of pyrimidine dimmers which inhibits the formation of new
DNA in the process of cell replication (Giese and Darby, 2000). These changes ultimately result in
mutations, damage to the genetic information, impairment of replication and gene transcription,
and then in the death of the microorganism cells. In photo-thermal effect, energy of the light pulses
is absorbed by the surface layers and is dissipated as heat, causing a certain rise in temperature
of about 130°C. This rise in temperature results in localized rapid heating of microorganisms
ultimately leading to rupture and cell death as short duration of light pulses prevents cooling of cell
surface by surrounding media (Wekhof, 2000).
Initial investigations on PLT, suggested microbial inactivation by PLT follow first-order kinetics
which was associated with only photochemical effect by UV-C; however, in further research
investigations survival curves showed variation from the linearity in the form of shoulders, tails, or
both. It was interpreted by the researchers that non-inactivation of microorganisms in shoulder phase
(lag-phase) is due to the presence of DNA repair system which is possible in low doses. Once this repair
capability is surpassed, the rate of inactivation increases even with little additional exposure to PLT.
After this, again the process becomes less efficient due to the tailing effect. The observance of tailing
effect is attributed to various factors such as existence of subpopulations with different resistance,
existence of cell aggregates, lack of a homogeneous dose distribution because of the high absorptivity
and/or presence of suspended solids in liquid foods, and the shading effect from surface irregularities
of solid matrix (Condon et al., 2014). The nonlinearity in inactivation behaviour of microorganisms
due to shoulder and tails are well explained by Weibullian models in several research investigations.
The effectiveness of PLT depends on process factors such as technical characteristics of the
lamp, type of quartz envelope of the lamp, electrical current density, voltage and pulse duration,
and food-related factors such as absorptivity in liquid media and the surface topography of solids.
Microorganisms express different tolerance levels towards PLT. The resistance of microorganisms
to PLT depends on type of cell; in general, gram-positive bacteria are more resistant to PLT than
gram-negative bacteria due to possible presence of more efficient DNA repair mechanisms in gram-
positive and variation in cell wall composition. Yeast eukaryotic cells demonstrate a little more
resistant than vegetative bacteria. Bacterial spores are much more resistant than vegetative cells
and fungal spores expressed highest resistance to PLT which requires intense treatments ultimately
leading to quality loss as an undesired side effect. The foremost drawbacks of PLT include poor
penetrating power of light and the requirements of transparent and smooth surfaces in the product
to be treated. Therefore, these limitations can be overcome by coupling different hurdles.
17.3.4.1 Pulsed Light Processing in Combination with Heat
Mild temperatures can be effectively used as postharvest treatment of fruits and vegetables
once coupled with PLT (Marquenie et al., 2002). The impact of the combining PLT and mild heat
treatments on microbial inactivation particularly bacterial spore reduction is not well addressed and
literature is scarce.
In a study, strawberries and sweet cherries were exposed to pulses of intense white light
(30 as at a frequency of 15 Hz) for duration varying from 1 to 250 s to inactivate conidia of the
fungi Botrytis cinerea and Monilia fructigena. As a result, maximal inactivation of 3 and 4 log
units for conidia of B. cinerea and M. fructigena, respectively, was observed with a similar
behaviour to pulsed light also, increase in inactivation rate was found with increasing treatment
intensity and complete inactivation was not achieved. Whereas, a synergism was obtained when
light treatment was combined with heat indicating a complementarily in lethal effects. Complete
inactivation of M. fructigena conidia was obtained after a 40 s pulsed light treatment and heating
for 15 min at 41°C, or after an 80 s light treatment and heating for 10 min at 41°C (Marquenie
et al., 2003).
Luz and Martínez de (2015) demonstrated that the combination of heat and PLT is a promising
preservation method. The combined preservation effect of PLT and heat processing on the
inactivation of Bacillus subtilis spores was assessed. B. subtilis spores subjected to sublethal pre-
treatments (PL or thermal treatments) were found to be more sensitive to subsequent PL or thermal
treatments than untreated spores. The order of treatment in which thermal treatment was followed
by PLT proves to be most effective combination in reducing B. subtilis spore counts. Bacterial
spores were found to continue in sensitized stage to successive treatment for at least 24 h of storage
in water (4°C or 30°C).
It is expected that the inactivation of bacterial spores in media of high absorptivity will be difficult
due to presence of small acid-soluble proteins (SASP) binding to the DNA; an efficient DNA repair
mechanism specific to SP; the accumulation of a high percentage of dipicolinic acid in the dormant
spore core; and the presence of a thick spore protein coating (Setlow, 2006). UV-C exposure to
bacterial spores results mainly in the formation of the “spore photoproduct” bipyrimidine dimers
(5-thyminyl-5,6-dihydrothymine) (Slieman and Nicholson, 2000).
17.3.4.2 Pulsed Light Processing in Combination with Antimicrobials
In response to consumer’s awareness about use of hazardous chemicals (such as chlorine)

for decontamination of fresh produce, alternative or complementary non-thermal processes
are being explored. In continuation with this, the potential of PLT in combination with natu-
ral antimicrobials to obtain minimally processed fruits and vegetables is investigated by few
researchers.
PLT has been approved by the FDA for treatment of foods with the total cumulative treatment
not exceeding 12.0 J/cm2 (FDA, 1996). Owing to reported outbreaks associated contamination of
imported green onion in the USA, efficacy of PLT coupled with surfactant and/or sanitizers to inac-
tivate Escherichia coli O157:H7 on stems and leaves of green onions inoculated by spot and dip
inoculation methods was studied. Result findings suggest that stems and dipped green onions were
more difficult to be decontaminated as compared to leaves and spot inoculated, respectively. The
combination of PLT and 1000 ppm of surfactant (sodium dodecyl sulphate) was found to be most
effective with highest degree of inactivation among tested treatments. This combination reduced the
E. coli O157:H7 population by 1.4 and 3.1 log10 CFU/g in dip inoculated stems and leaves of green
onions, respectively, stating it as a potential combination for decontamination (Xu et al., 2013).
Another combination of PLT and 10 ppm of sodium dodecyl sulphate was found to be most effective
for inactivation of Salmonella on green onions (Xu et al., 2015).
Combining PLT with antimicrobials (nisin or natamycin) to overcome the limitation of low
penetration ability associated with PLT was explored to achieve microbial inactivation in cheese.
An important finding of this investigation states that the order of treatments is critical to achieve
desired objective. Treatment with antimicrobials (nisin or natamycin) prior to PLT decreased the
effectiveness of PLT for all tested bacteria. While, increased inactivation was obtained when
antimicrobials were applied after PLT. A maximum reduction of 3.73 ± 0.96 log of L. innocua
was obtained at 9.22 J/cm2 for PLT followed by nisin, compared with 3.01 ± 0.48 by PLT alone
suggesting its prospective application (Proulx et al., 2017).
PLT in conjugation with antimicrobial edible coating (modified chitosan containing a
nano-emulsion of mandarin essential oil) was assessed for its efficacy against Listeria innocua
inoculated on green bean samples. A slight antagonistic effect on inactivation rate along with a
slight detrimental impact on colour properties was reported, while, the combination of the coating
application with HHP showed synergism of antimicrobial effects (Francesco et al., 2015). The
impact of PLT, alginate coating (ALC) and malic acid (MA) dipping treatments on fresh-cut mango
inoculated with L. innocua was assessed. Various combinations of PLT, ALC, and MA were
attempted to examine possible synergisms among treatments. Results revealed an additive effect
of MA-PLT and PLT-ALC-MA (the order of treatment being the critical parameter) combination
with reduction of L. innocua counts by 4.5 and 3.9 logs, respectively. The antimicrobial effects of
MA have been reported to be associated ionization of undissociated molecules and the resultant
decrease in intracellular pH (Rathnayaka, 2013; Ramos-Villarroel et al., 2014). An optimal
combination with optimal order of different treatments enables to ensure safety (Salinas-Roca
et al., 2016).
17.3.5 pulsed Electric Field in Combination with

PEF technology employs the use of short pulses of high electric field (20–80 kV/cm) for a
very short duration which varied from microseconds to milliseconds (Amiali and Ngadi, 2012).
Food processed by PEF maintains the natural-like or fresh characteristics with minimal loss of
the quality. PEF has a great potential as an alternative to traditional heat processing and is widely
accepted for processing of liquid, semi-liquid, and to some extent of solid food (Ho and Mittal,
2000; Jaeger et al., 2014; Yogesh, 2016). The bactericidal effectiveness of PEF depends on process
parameters (electric field intensity, power, and treatment time), microbial parameters (types, size
and shape, and growth phase), and on composition of food product to be processed (Ngadi et al.,
2009; Raso et al., 2014). The application of high-intensity pulses cause electroporation in the cell
wall of microorganism and are responsible for antimicrobial effects.
The mechanism of microorganism inactivation by PEF treatment involves three phases:
Phase I: There is a creation of pores in the cell membrane when PEF is applied for a short
duration of few nanoseconds to milliseconds. Phase II: The number and size of pores created
by PEF increases with the duration of nano to milliseconds, and Phase III: This is post-PEF
application phase wherein the cell death or complete inactivation occurs and the duration lasts
from milliseconds to few hours, or in case of incomplete inactivation cells start their resealing.
In such cases, the damage to microbial cell is sub-lethal and PEF-treated cells regain their viable
state.
PEF expressed significantly high effectiveness towards inactivation of vegetative cells of bacteria,
yeast, and moulds (Wouters and Smelt, 1997). However, PEF effectiveness towards inactivation
of bacterial spores and enzymes articulated mixed results. Some authors reported insufficiency
of PEF in inactivating bacterial spores (Grahl and Markl, 1996; Pagan et al., 1998) while others
have reported the inactivation of bacterial spores (Marquez et al., 1997) and enzymes by PEF
(Ho et al., 1997; Giner et al., 2000). Depending on the process parameters and food composition,
electroporation can be reversible or irreversible, in irreversible electroporation there is a permanent

damage to membrane while reversible damage has the ability to reseal the pores indicating sub-
lethal injuries and requirement of combination processing technologies. Consequently, it is well
established by several researchers that PEF when used alone possess limited bactericidal effects;
hence, its use in combination with other conventional and non-conventional barrier technologies is
recommended (Bermudez-Aguirre et al., 2012; Siemer et al., 2014).
17.3.5.1 Pulsed Electric Field in Combination with Heat
In view of above recommendation, one possible combination is use of PEF with moderate
heat treatment. The advantage of coupling moderate PEF and moderate heat is explained by
the fact that higher temperatures induce structural changes leaving the cell membranes more
susceptible to PEF-induced electroporation, resulting in increased efficacy or synergistic effect
(Heinz et al., 2003; Bazhal et al., 2006). The plausible benefit of this synergistic lethal effect is
the likelihood of reducing the intensity of the PEF treatment and hence, its adverse effects in
achieving the goal of food safety. Jin et al. (2009) studied the effects of PEF and temperature
on S. typhimurium DT104 in liquid whole egg and suggested a 3-log10 reduction after PEF treat-
ment (25 kV/cm) for 250 μs at 55°C which was comparable to that of thermal treatment at 60°C
for 3.5 min. Zhao et al. (2007) indicated that PEF with a bipolar pulse (2 µs wide), an intensity
of (30 kV/cm, 100 Hz) with the processing temperature of 40°C and treatment time for 800 µs,
was sufficient to achieve pasteurization conditions with respect to S. enteritidis, E. coli and
S. aureus as common spoilage and pathogenic micro-organisms in egg products. The effects of
inlet temperature (20°C–45°C), electric field strength (EFS; 20–42.5 kV/cm) and treatment time
(68–170 μs) on microbial inactivation by PEF was studied by Cregenzán-Alberti et al. (2015)
using a central composite design. Results expressed reductions of 5 and 5.2 log10 cycles in popu-
lation of E. coli and S. aureus, respectively, at 32.5°C, 40 kV/cm and 89 μs while, P. fluorescens
was reduced by 5.3 log10 cycles at 32.5°C, 42.5 kV/cm and 106 μs. Similar trend was found by
Dutreux et al. (2000) with a PEF treatment of 41 kV/cm for 157.5 μs at 37°C in fat-free milk
resulting in a 4-log10 cycle reduction of E. coli.
17.3.5.2 Pulsed Electric Field in Combination with Acidic pH
The role of pH in the survival of microorganisms is associated with their ability to maintain the
cytoplasmic pH near neutrality, therefore, pH above or below neutrality decreases the resistance of
microorganisms and inhibits the germination of spores. This effect is well adapted in hurdle con-
cept while using thermal processing, therefore, this effect can be explored for its potential in hurdle
approach while utilizing non-thermal processing technologies. PEF in combination with low pH
has provided varied degree (synergistic and antagonistic effect) of sensitivity towards inactivation
of vegetative bacteria cells.
García et al. (2005a) reported varied degree of inactivation by PEF and pH combination,
depending on the type of bacterial cell, at acidic pH, the resistance gram-positive bacteria towards
PEF declines, while gram-negative bacteria were more PEF resistant. The greater PEF resistance
of gram-positive bacteria at neutral pH might be attributed to the reversible damage (expressing the
possibility to repair membrane damage) which would become irreversible at acidic pH rendering
the cells more sensitive to PEF. In contrast, the higher PEF resistance of gram-negative bacteria at
acidic pH would be correlated to the ability to repair the cytoplasmic membrane when extensively
damaged. The bactericidal effects of PEF (12.5 kV/cm, 1 pulse) on E. coli O157:H7 in a 10%
glycerol solution showed synergism when pH of the medium was lowered from 6.4 to 3.4 using
in the presence of 1,000 ppm of either sodium benzoate or potassium sorbate (Liu et al., 1997).
This synergistic effect is associated with the fact that non-dissociated fraction of organic acids
has the ability to penetrate the bacterial cell wall and disrupt its pH homeostasis and efficacy
of this combination depends on the ratio of the dissociated and non-dissociated portions of the
organic acid. In contrast to this, PEF in combination with acidification by using hydrochloric acid
resulted in no additional reduction in raw milk’s microflora when compared with PEF alone (Smith
et al., 2002). The effect of pH reduction (from 6.30–6.45 to 4.22–4.46) by the addition of sodium
benzoate and potassium sorbate and PEF at 13–15 µs/25–50 Hz on the inhibition of Saccharomyces
cerevisiae in prickly pear beverages resulted in a synergistic effect by achieving reduction of at
least 3–4 log10 immediately after processing, and more than 5 log10 on fourth day of storage at 25°C
(García-García et al., 2015). Due to mixed results expressed by this combination, extensive research
is required before these technologies can be adapted at commercial level production of shelf-stable
low-acid foods.
17.3.5.3 Pulsed Electric Field in Combination with Antimicrobial
Combining antimicrobials with PEF suggested a plausible hurdle combination due to

enhanced inactivation effects in achieving product stability and safety. Antimicrobials such as
EOs are attracting researcher’s interest for their potential as barrier in combination with non-
thermal technologies, as they possess “generally recognized as safe” (GRAS) status and global
acceptance from consumers. Synergistic effects of PEF coupled with cinnamon bark oil at 0.05%
against S. enteritidis and L. monocytogenes and for E. coli O157:H7 at 0.1% in melon and water-
melon juices was reported by Mosqueda-Melgar et al. (2008). In another study, combination of
PEF with 1.3 mM carvacrol showed synergism with reduction of more than 5 log10 cycles after the
application of 20–50 pulses depending on the juice type (apple, mango, orange, and tomato juice
(Ait-Ouazzou et al., 2013).
Among bacteriocins, nisin is the most widely studied bacteriocin in combination treatments
of non-thermal processing technologies due to its GRAS status. Nisin causes ion-permeable pore
formation in the cytoplasmic membrane of gram-positive bacterial cells. Synergistic effect was
obtained by combination treatment of PEF (100 μs at 40 kV/cm) and nisin (2.5 IU/mL) in orange
juice against survival of Listeria innocua with additional reduction of 1.7 log than the sum of PEF
and nisin independent effects (McNamee et al., 2010). A similar synergistic effect of combination
of PEF (80 kV/cm, 20 pulses, 50°C) and nisin (100 IU/mL) was reported by Nguyen and Mittal
(2007) on natural microflora of tomato juice by achieving an additional 2.2 log cycles reduction in
comparison to the sum of the individual effects.
17.3.6 Ozone in Combination with Conventional preservation technologies
Ozone has been recognized as safe (GRAS) and is approved by United States Food and
Drug Administration for use as a disinfectant or sanitizer in foods and food processing in US
(USDA, 1997). US FDA granted GRAS status for use of ozone in bottled water in 1982 and
for the treatment, storage, and processing of foods in 2001 (Khadre et al., 2001). Ozone is a
strong oxidant and that owes to its bactericidal properties. The antibacterial effect of ozone
involve oxidation of sulfhydryl groups and amino acids of enzymes, peptides, and proteins
to shorter chain to peptides and oxidation of polyunsaturated fatty acids to peroxides which
results in cell disruption and leakage of cellular components (Guzel-Seydim et al., 2004a). The
efficacy of ozone depends on the type of food matrix to be treated, smooth surfaces are easy to
decontaminate than foods with rough or uneven surface. The organic material in food product
such as proteins and fats, significantly diminishes the efficacy of ozone as these components
may compete with contaminating microorganisms for the ozone (Guzel-Seydim et al., 2004b).
Therefore, in order to improve efficacy of ozone, combination of ozonation with other technolo-
gies which can either support in generation of free radical or can reduce the uptake of ozone
by organic components, is recommended. In this line, the combination of ozone with oxidant
such as hydrogen peroxide, heat, non-thermal technologies namely UV, ultrasound and PEF has
been addressed by researchers.
17.3.6.1 Ozone in Combination with Heat
Radicals generated during ozone decomposition possess better and faster bactericidal properties
due to their higher reaction rates as compared to ozone molecules themselves. Kim et al. (2003) did
not suggest simultaneous application of heat and ozone in combination processing as it is ineffective
against food microflora due to rapid degradation of ozone with heat; however, sequential application
of ozone and heat may prove to be beneficial while, Sung et al. (2014) reported a synergistic effect
in the inactivation of pathogens in apple juice treated with ozone and heat simultaneously. They
investigated the combination effect of ozone and heat treatments in apple juice for the inactivation
of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes. Apple juice
inoculated with these pathogens were simultaneously exposed to gaseous ozone (2.0–3.0 g/m3) and
heat (25°C, 45°C, 50°C, and 55°C) for up to 1 min. A decline in surviving populations with increase
in temperature was attained. Combination treatment of ozone and heat for 1 min reduced E. coli
O157:H7 by 1.50 and 1.60 log CFU/mL, respectively, at 25°C and 45°C, and below the detection
limit (1 log CFU/mL) at 50°C and 55°C. A similar trend was observed with S. typhimurium and
L. monocytogenes and a synergistic effect was observed at 50°C.
The antimicrobial efficacy of gaseous ozone in combination with heat (4°C, 20°C, and 50°C)
was assessed by Williams et al. (2004) in unpasteurized apple cider and orange juice containing
E. coli O157:H7 and S. typhimurium. The bactericidal effects were expressed in following order:
50°C > 4°C > 20°C. Heat treatment at 50°C after 45 min of ozone treatment resulted in inactivation
of >5.0 log CFU/mL while less than 5.0 log CFU/mL reduction was obtained after treatment at 20°C
for 3–4 h.
17.3.6.2 Ozone in Combination with Antimicrobials
At high pH, the rate of ozone decomposition increases due to the reaction of free radicals in
aqueous ozone with hydroxyl ions; therefore, a low pH is the requirement for increased efficiency of
ozone that can be achieved by adding organic acids which in turn results in enhanced efficacy due
to additive or synergistic effect. The microbial inactivation mechanism by organic acids has been
linked with undissociated acid molecules that impede with cellular metabolism or a decrease in
biological activity associated with change in pH of cell’s environment. The combined treatment of
ozone and organic acid (acetic, citric, or lactic acids) was more effective than individual treatments
in reducing initial population levels of Escherichia coli O157:H7 and Listeria monocytogenes on
enoki mushroom (Yuk et al., 2007) and lettuce (Yuk et al., 2006). A reduction in postharvest decay
and pericarp browning of longan fruit when treated with combination of ozone and citric or oxalic
was reported by Whangchai et al. (2006).
The reaction between ozone and hydrogen peroxide when combined in aqueous solution
generates hydroxyl free radicals. Being a weak acid, hydrogen peroxide partially dissociates
into hydroperoxide ion in aqueous solutions and this hydroperoxide anion is highly reactive
with ozone as compared to hydrogen peroxide molecule. However, the increased pH due to
hydroxyl radicals results in ozone decomposition therefore, appropriate concentration of H 2O2
is required to avoid adverse reactions and to achieve antimicrobial effect. Williams et al. (2005)
demonstrated that combining ozone and dimethyl dicarbonate or hydrogen peroxide followed
by refrigerated storage for 24 h can be used to achieve a 5-log CFU/mL reduction of E. coli
O157:H7 and Salmonella in apple cider and orange juice indicating continued lethality of
antimicrobials during refrigerated storage.
17.3.7 Electron Beam processing in Combination with

Microbial inactivation by electron beam processing (EBP) follows direct or indirect routes, lead-
ing to cell injury or death. Direct route involves, the damage of the physiological metabolism due to
the generation of energy transfer within the body of microorganisms, consequently the destruction
of chemical and molecular bonds (breaks in DNA structure, and denaturation of enzymes and mem-
brane proteins) hindering the normal physiological metabolism activities and cell replication which
ultimately leads to cell death (Miller, 2005). The indirect route follows generation of unstable free
radicals by ionization of water molecules on exposure to e-beam, resulting in intracellular oxidation
progressing to cell injury and death. The effectiveness of decontamination effect of the e-beam on
processing factors such as irradiation dose (generally directly proportional to the degree of killing
microbes), food-related factors (food composition), and microorganism-related factors (depends on
the species of targeted microorganisms). Some types of microorganisms are known to survive low-
dose irradiation, and therefore high-dose irradiation is required to kill these organisms which pose
possible damage to the sensory characteristics and nutritional quality of food. When the inherent
or added factors prevailing food preservation such as density, temperature, pH, and composition of
gases are adverse to microbial growth, their resistance to the e-beam decline resulting in killing
at lower dose of irradiation. Therefore, careful selection in combining e-beam with other possible
conventional or non-thermal hurdles is recommended. Microorganisms illustrate different degrees
of tolerance intensity towards EBP. In general, prokaryotes are supposed to be more resistant than
eukaryotes, gram-positive bacteria exhibit greater degree of resistance than gram-negative bacteria
(Moosekian et al., 2012).
17.3.7.1 Electron Beam Processing in Combination with Antimicrobials
The hurdle effect of electron beam irradiation (2 kGy) in amalgamation with citrus peel extract
(2%) resulted in complete inactivation of viable cells in marinated chicken breast meat, while
electron beam alone at 2 kGy reduced the initial microbial counts by 3.16 log cfu/g. Although, this
combination proved to be successful in achieving the objective of shelf life extension of marinated
chicken breast but adversely affected the sensory characteristics, therefore, further research efforts
are required to avert undesirable effects (Alahakoon et al., 2015). A similar trend was observed
when electron-beam processing used in combination with leek (Allium tuberosum Rottler) extract
on the quality of pork jerky during ambient storage, a notable reduction in total aerobic bacterial
count with an increase in the irradiation dose and leek extract addition in comparison to that of the
control was achieved however, a decrease in sensory flavour and overall acceptability was resulted.
Therefore, in order to meet market requirements, suitable combinations in hurdle approach with
simultaneous retention of sensory quality are warranted (Kim et al., 2013). A study, where EBP
(4 kGy) was mixed with onion peel extracts and barbecue flavouring (0.5%), elucidate no viable
cells while maintaining the sensory quality in terms overall acceptability as compared to con-
trol sample of pork jerky (Kim et al., 2012). Therefore, such combinations which can effectively
encounter both the objectives of achieving food safety and maintaining sensory quality are the
need of the hour.
17.4 hUrDLE apprOaCh IN COMBINING NON-thErMaL tEChNOLOGIES
Few attempts have been made at laboratory scale for combining non-thermal technologies in var-
ious food products and different degree of results was obtained. Therefore, this area needs extensive
research before its commercial application. Some of the invested combinations are discussed here.
17.4.1 high hydrostatic pressure and Ozone
The mechanism by which ozone kills vegetative bacterial population is known to be associated
by reaction of ozone with microbial cell membranes which is believed to cause the oxidation of lip-
ids present on the bacterial cell envelope which eventually directed to leakage of intracellular cell
contents and damage of genetic material leading to cell death (Kim et al., 2003). Bacterial spores
are the most resistant to ozone and bacterial vegetative cells are the most sensitive, stationary-phase
cells appears to be more resistant than are cells from the exponential phase. However, due to struc-
tural differences between the vegetative and dormant states of a bacterial cell, the mechanism of
spore destruction by ozone is not well understood.
A study was conducted to assess the ability of a combined hurdle technology of aqueous ozone
and high-pressure processing to inactivate superdormant spore (which germinate extremely slowly)
populations of mesophilic and psychro-tolerant isolates of Bacillus cereus. The results established
that superdormant spores were approximately 20% more resistant to ozone treatment than hetero-
geneous spore populations and psychro-tolerant species were approximately 31.9% more resistant
than mesophilic species. A maximum reduction of 2.67 log CFU/mL was achieved in superdormant
spores when treated with a combination of ozone-HHP hurdle technology (Markland et al., 2013).
17.4.2 high hydrostatic pressure and Microfiltration
Microfiltration (MF) employs the use ceramic membranes (pore size of 1.4 mm and in a cross-
flow setup) to reduce the microbial load without affecting the native state of proteins. Approximately
above 3.5 log cycles reduction in microbial load is reported to be achieved by MF (Saboyainsta and
Maubois, 2000; Fritsch and Moraru, 2008; Walkling-Ribeiro et al., 2011). Colostrum, a bovine
secretion is a unique source for biomolecules and a valuable raw material for the nutraceuticals
production. The heat-sensitive nature and unpredictability in microbial quality of raw colostrum
possess difficulty in manufacture of safe and stable colostrum products for human use. A novel
combination of MF and subsequent application of high pressure was explored. Skimmed colostrum
contaminated with various bacterial species, subjected to cross-flow MF (pore sizes of 1.4 and
0.8 mm) with subsequent exposure to HHP at 400 and 500 MPa for 10 min, resulted in reduction of
residual microbial burden to undetectable levels in MF permeates (Gosch et al., 2014).
17.4.3 high hydrostatic pressure and Carbon dioxide
The inactivation of spores by high-pressure CO2 follows a two-step pathway indicating penetra-
tion of CO2 into the cells and subsequent destruction and inactivation of germinated cells (Setlow
et al., 2016).
Alicyclobacillus acidoterrestris, a thermoacidophilic spore-forming bacterium is a spoilage
causing agent that can develop post-pasteurization in juices. The combined effect of innovative
technologies such as HHP and supercritical carbon dioxide (SCCD) was evaluated for inactivation
and germination of spores of two A. acidoterrestris strains, suspended in apple juice. The results
indicated a significantly high resistance of spores of the A. acidoterrestris TO-117/02 strain towards
SCCD and HHP on the used parameters and further demonstrate that SCCD and HHP combined
with moderately elevated temperature maybe a useful technique for inactivation of A. acidoterrestris
spores in single strength juices (Izabela et al., 2017).
The combined outcome of HHP and dense-phase carbon-dioxide process on inactivation of
enzymes (POD, PPO, and PME) in feijoa (Acca sellowiana) puree was investigated. Reduction in
enzyme activities of tested enzymes was observed with increasing treatment time, while increasing
CO2 level decreased PME activity, and pressure influenced PPO activity. The lower pH values
also pose an additional advantage in increasing enzyme inactivation. The synergism was obtained
at optimal process conditions of 13 min, 600 MPa, pH-3, CO2 saturation, with resultant residual
activity of 74.3% ± 3.3%, 70.9% ± 2.6%, and 53.9% ± 0.9% for POD, PPO and PME, respectively
(Duong and Balaban, 2014).
17.4.4 high hydrostatic pressure and Gamma Irradiation
The exposure of gamma irradiation (1.0 kGy) and HHP (200 MPa for 30 min) alone or in combina-
tion on the shelf-life of lamb minced meat at 0°C–3°C with initial counts of 105 CFU/g total microbial
load, 102 CFU/g of coliforms and 104 CFU/g of Staphylococcus spp. resulted in complete elimination
of coliforms while, Staphylococcus spp. however, were reduced only by 1 log cycle when treated with
irradiation alone and high pressure alone. Amalgamation of gamma irradiation and HHP displayed
appearance of Staphylococcus spp. only after 3 weeks of storage. On the basis of microbiological
and sensory quality, control sample expressed the shelf-life of less than 1 week while, all treatments
extended shelf-life of meat samples by 3 weeks however, only combination treatment resulted in meat
samples free from potentially pathogenic Staphylococcus spp. (Paul et al., 1997).
17.4.5 Ultrasonication and Supercritical Carbon Dioxide
Supercritical carbon dioxide (SC-CO2) has been increasingly explored for its potential to stim-
ulate the inactivation of the natural microbial flora as well as pathogens. CO2 used in the process
has a GRAS status with additional advantages of being relatively inert, inexpensive, nontoxic,
nonflammable, recyclable and ready availability in high purity and no residues after use. The criti-
cal temperature (31.1°C) is suitable for the applications where thermal stability is important, and
the critical pressure (7.3 MPa) is easily achievable in industrial processes. Possible mechanisms
of microbial inactivation by SC-CO2 involve the diffusion and solubility of SC-CO2 in the culture
medium, the resultant decrease of the pH in the medium, the increase of the membrane fluidity and
permeability, the diffusion of CO2 into the cells, the cell membrane rupture caused by the increase
of the internal pressure, and the resultant changes in the cellular environment, such as a decrease in
pH, the inactivation of key enzymes, and the extraction of critical intracellular materials (Garcia-
Gonzalez et al., 2007). Due to limitations associated with SC-CO2 process, combination with other
preservation techniques using hurdle approach is recommended.
A study was conducted to witness the effect of SC-CO2 alone or in combination with HPU on
natural microbial flora and inoculated Salmonella enteric in coconut water. The synergistic effect of
SC-CO2 and HPU was evident with a finding that 15 min is required to achieve about 5-log reduction
in combination, while it took 30 min when SC-CO2 was used alone. Also, marked effect was
observed during storage study which highlighted heavy regrowth during the storage period in case
of SC-CO2-treated coconut water, while, a full shelf life of 4 weeks was guaranteed for SC-CO2 and
HPU combination (Cappelletti et al., 2014). Sara et al. (2014) demonstrated that HPU alone was not
able to induce any microbial inactivation in dry-cured ham spiked with Listeria monocytogenes
with an initial concentration of about 109 CFU/g while combination of high-pressure CO2 and HPU
process assured inactivation to undetectable level with conditions of 12 MPa, 35°C, at 10 W for 5 min
and a full shelf-life of 4 weeks at 4°C was guaranteed. Whereas, the combined treatment was not able
to induce a greater reduction of the natural microbial flora at milder process conditions compared to
high-pressure carbon dioxide alone in case of fresh-cut carrot (Giovanna and Sara, 2015).
17.4.6 Ultrasonication and Osmotic pressure (Osmo-sonication)
High osmotic pressure is apparently known to cause metabolic perturbation by inhibiting the
glucose phosphotransferase system, a mechanism by which microorganisms take up glucose (Walter
et al., 1987). In natural juices, neither sonication nor high osmotic pressure alone adequately achieves
microbial inactivation. Therefore, the efficacy of US (50 ± 0.2 W, 20 kHz) to reduce Salmonella
in conjugation with subsequent concentration and storage at high osmotic pressure was evaluated
in different solutions (PBS, sucrose, and orange juice) by Wong et al. (2012). Sonication alone did
not cause significant membrane damage while storage at high osmotic pressure (10.9 MPa) for 48 h
only affected membrane permeability in 20% of cells. When the contaminated orange juice was
subjected to combined treatment, a 5-log10 cfu/mL reduction of Salmonella spp. was achieved. In
view of synergistic effect of combination, authors suggested that osmo-sonication is an innovative
alternative for the non-thermal decontamination of liquid foods.
17.4.7 Ultrasonication and Ultraviolet radiation (photosonication)
The lack of penetrating efficiency of UV radiation (UV-C radiation, germicidal UV at 254 nm)
in turbid and coloured liquids (Sizer and Balasubramaniam, 1999) necessitate to combine with
such a treatment which can improve the penetrating power and hence the lethality of UV radiation.
Furthermore, many nutrients (vitamins, pigments, fatty acids, etc.) are sensitive to UV light; there-
fore, low doses or combination technologies are preferred to prevent undesirable effect along with
desired microbial destruction.
Ultrasound has been thought of one such kind of technology which can enhance the efficiency
of UV light penetration and lethality against microbial destruction due to the turbulence created
by cavitation (formation and implosion of bubbles) in a liquid media. The raw milk was subjected
to acoustic energy and UV irradiation (photosonication) simultaneously in order to reduce total
and coliform bacteria, a reduction of 4.79 log cfu/mL and 5.31 log cfu/mL for total and coliform
bacteria was achieved as compared to 1.31 log cfu/mL and 4.01 log cfu/mL in sonication alone.
This enhanced effect was attributed to the complementary effect of both the technologies on each
other. Furthermore, combination of ultrasound and UV light was found to be more efficient with
respect to treatment time and energy consumption compared to either treatment individually
(Sengulf et al., 2011).
The combined effects of decontamination efficacy of UV light and ultrasound on freshly
cut lettuce and strawberry inoculated with a cocktail of four bacteria (Escherichia coli, Listeria
innocua, Salmonella enteritidis and Staphylococcus aureus) were investigated. Treatment with
UV and US reduced the population of selected inoculated bacteria on lettuce and strawberries
without adversely affecting the colour of produce. Consequently, this combination could prove to be
a promising, non-thermal and environmentally friendly alternative to chemical disinfectants such as
chlorine and hydrogen peroxide solutions for fresh produce industry (Birmpa et al., 2013).
17.4.8 Ultrasonication and pulsed Light processing
The destructive effect of high-intensity light pulses (HILP) on microorganisms is typically due
to the photochemical action of the UV-C part of the light spectrum directly on DNA. This effect is
responsible for thymine dimerization in DNA which prevents cell replication and ultimately leading
to death (Gómez-López et al., 2007).
The application of non-thermal technologies such as HILP and TS in combination at two
different energy settings (Low (L) and High (H)) to inactivate Escherichia coli in orange juice
revealed inactivation ranging from 1.10 (TS(H)) to 2.42 (HILP(H)) log cfu/mL for the hurdles
when applied individually and from 2.5 (HILP(L)&TS(H)) to 3.93 (HILP(H)&TS(L)) log cfu/mL
for the combined treatments (Muñoz et al., 2011). The effect of ultrasound (US) and PLT on the
inactivation of Alicyclobacillus acidoterrestris ATCC 49025 spores and Saccharomyces cerevisiae
KE162 inoculated in commercial and natural squeezed apple juices was evaluated. The results
interpreted that inactivation rate was dependent on treatment time, temperature, microorganism,
and matrix. Combination of these technologies directed reduction of up to 3.0 and 2.0 log cycles of
spore in commercial and natural apple juice, respectively; while, this reduction was 6.4 and 5.8 log
cycles in commercial and natural apple juices, respectively for S. cerevisiae. In natural apple juice,
the combination of US—PLT at the highest temperature build-up of 56°C ± 1°C was the most
effective treatment for both the strains. It was established that certain combinations of US and PLT
kept on good microbial stability under refrigerated conditions for 15 days (Ferrario et al., 2015).
Therefore, optimal combinations required to be established for better outcomes in terms of food
safety.
17.4.9 Ultrasonication and Ionizing radiation processing
Ionizing radiation in combination with US was evaluated for its effectiveness in reducing
Bacillus cereus F4810/72 spores in raw rice. A higher degree of destruction was achieved in com-
bination in comparison to individual treatments alone. The combined treatment of 0.1, 0.2, and
0.3 kGy and US (5–20 min) completely destroyed the spores in raw rice (Ha et al., 2012).
17.4.10 Ultrasonication and pulsed Electric Field processing
Ultrasound when applied in the form of TS or MTS with PEF articulated fruitful results. TS
and PEFs represent commercially viable, emerging technologies for preservation of liquid food.
Orange juice was treated with a combination of TS and PEF to achieve pasteurization effect. The
exposure of juice to batch TS at 55°C for 10 min followed by continuous PEF at a field strength of
40 kV/cm for 150 s resulted in overall microbial counts within safe levels (<1000 CFU/mL) dur-
ing the 168 days of storage. However, the counts were higher than in thermally pasteurized juice
(94°C for 26 s) (Walkling-Ribeiro et al., 2009). The combination of TS and PEF was tested for its
efficacy against survival of Listeria innocua 11288 (NCTC) in milk. The tested maximum field
strength (40 kV/cm) combined with TS (80 s) led to 6.8 log10 CFU/mL inactivation while, milk
pre-heated to 55°C (over 60 s) prior to TS followed by PEF (30 and 40 kV/cm) showed inactivation
between 4.5 and 6.9 log10 CFU/mL which is comparable to thermal pasteurisation. The research
findings signify that TS followed by PEF represents a valid alternative for L. innocua inactivation
in milk (Noci et al., 2009).
17.4.11 Ultrasonication and high hydrostatic pressure (Mano-sonication)
The combination of ultrasound and pressure at sub-lethal temperatures is known as mano-

sonication (MS) and is reflected to present an unconventional approach to boost cavitation effects
in liquid media. When high pressure is applied externally, the pressure increases the cavitation to
a higher threshold resulting in more violent and rapid bubble collapse, hence, better inactivation
results. In a study on inactivation of L. monocytogenes by high-power ultrasonic waves (20 kHz,
117 mm) at ambient temperature in combination with pressure, a decline in D value to 1.5 min
at 200 kPa and 1.0 min at 400 kPa was achieved as compared to 4.3 min when ultrasonic waves
were used alone. An exponential increase in inactivation rate by MS with the ultrasonic wave’s
amplitude in the range of 62–150 mm was reported. An amplitude increase of 100 mm decreased
the MS resistance of approximately six times and no effect of by treatment temperature up to
50°C was observed (Pagan et al., 1999). Fresh apple juice was subjected to US treatment (25 kHz
and 70% amplitude) at 20°C for 60 min followed by HHP treatment at 250, 350, and 450 MPa
for 10 min at room temperature, revealed that the combined US-HHP at 450 MPa resulted in
complete inactivation of total plate counts, yeasts, and moulds with highest degree of enzyme
inactivation. The results recommend that the combination of US and HHP can act as a potential
hurdle to produce safe and high-quality apple juice with reduced enzymes and microbial activity
and improved nutrition (Abid et al., 2014).
17.4.12 Ultrasonication, heat and high hydrostatic

pressure (Mano-thermo-sonication)
The combination of ultrasound and heat under pressure is known as MTS. The prime benefit of
this combination is that the loss of the cavitational effect at temperatures near the boiling point due
to the high–water vapor pressure in the liquid medium can be overcome by applying an external
pressure. As a result, cavitation can also be achieved at higher temperatures (above the water boiling
point); therefore, enhanced inactivation effects can be achieved. This combination has been used
effectively for destruction of spore-forming bacteria (Earnshaw et al., 1995).
During MTS treatment, the cells are subjected to intense vibration and cavitation, and undergo
a violent shaking which results in release of DPA and low-molecular-weight polypeptides from
spores (Palacios et al., 1991). Also, degradation of the cortex is reported to cause the rehydration
of protoplast which in turn results in loss of heat resistance, making the cells more susceptible to
the heat and ultrasound treatment. A decline in D78°C-value (at 20.2 W/mL) from 28 to 14 min was
reported when the orange juice was previously submitted to 600 MPa for 15 min. Authors concluded
that pressure-assisted TS provided the best method for spore inactivation with favourable benefits of
all three hurdles (Evelyn and Silva, 2016).
The effectiveness of sonication, MS, TS, and MTS treatments were compared with controlled
thermal treatment at the same temperatures in response to inactivation of Escherichia coli K12.
Negligible inactivation was obtained at 40°C, 47°C, and 54°C, while at 61°C a 5-log reduction was
achieved. In sonication alone at sub-lethal temperatures (40°C, 47°C, and 54°C) a 4-log reduction
was attained. On the other hand, MS at sub-lethal temperatures resulted in 5-log reduction at about
2 min. Furthermore, at 61°C, the treatments TS and MTS further increased the inactivation by
achieving 5 log reduction in only 0.5 min. Also, it was evident that MTS and TS resulted in almost
identical inactivation rate, especially in the first 0.75 min of the treatment (Lee et al., 2009). The
results clearly signify the advantages of TS and MTS combinations over others.
MTS combination also proves to be efficient for inactivation of various enzymes such as
lipoxygenase, POD, PPO, proteases, and lipases from psychrotrophic bacteria (Lopez et al., 1994;
Sala et al., 1995; Vercet et al., 1997). The simultaneous application of heat (72°C) and ultrasound
under moderate pressure (200 kPa), increases the inactivation rate of heat-resistant PME 25 times in
buffer and >400 times in orange juice (Vercet et al., 1999).
17.4.13 pulsed Electric Field and high-pressure Carbon dioxide
PEF and high-pressure CO2 is one of the successful combinations among non-thermal process-
ing technologies. Spilimbergo et al. (2014) reported synergistic interaction between PEF and high-
pressure CO2 for inactivation of Salmonella enterica typhimurium cells in 10% glycerol solution.
The subsequent application of PEF for 1 ms, 1 pulse, 30 kV/cm and 4 ms, 12 pulses, 30 kV/cm
and high-pressure CO2 at 22°C, 12 MPa from 0 to 45 min depicted decrease in viability of bacte-
rial cells as observed by flow cytometry. This combination when investigated against survival of
Escherichia coli, Staphylococcus aureus, and Bacillus cereus in glycerol solution, synergistic effect
was observed in sequential application of PEF (up to 25 KV/cm) and then with high-pressure CO2
(up to 40°C and 200 bar) as determined by scanning electron microscopy. Also, a decline in cell
viability with increasing electrical field strength and number of pulses was observed. Furthermore,
a batch treatment with supercritical CO2 lead to complete inactivation of bacterial species and
decreased the count of the spores by at least three orders of magnitude due to increased contact time
between CO2 and the sample (Spilimbergo et al., 2003). The synergistic effect of this combination is
attributed to the partial structural damage which supported the subsequent CO2 penetration into the
cell and increasing the inactivation kinetics in addition to electroporation effect of PEF. Although
the combination expressed positive results, extensive studies are required to validate the viability
of this non-thermal pasteurization process at laboratory and industrial level in real food matrices as
they possess more compositional complexity.
17.4.14 pulsed Electric Field and Ozone
PEF in conjugation with ozone was explored for its potential synergy against food-borne bac-
teria such as Lactobacillus leichmannii ATCC 4797, Escherichia coli O157:H7 ATCC 35150, and
Listeria monocytogenes Scott A by Unal et al. (2001) when suspended in 0.1% NaCl. The results
expressed varied degree of synergy among different treatment levels and the bacterium treated,
with reduction of 7.2 log10 CFU/mL in L. leichmannii cells (Ozone exposure; 1.00 µg/mL of
ozone followed by PEF; 20 kV/cm), 3.6 log10 CFU/mL in E. Coli (Ozone; 0.75 µg/mL followed by
PEF; 15 kV/cm) and 3.9 log10 CFU/mL in Listeria monocytogenes (Ozone exposure; 0.75 µg/mL of
ozone followed by PEF; 15 kV/cm). Combination of ozone, UV, and PEF yielded additive bacteri-
cidal effect in population of Escherichia coli 0157:H7 in poultry chiller water (Ngadi et al., 2004).
17.4.15 pulsed Electric Field and Microfiltration
Sequential combination of PEF and MF represents a potential alternative for “cold” pasteurisa-
tion of milk with improved quality characteristics. Combination of PEF followed by MF obtained
equivalent reductions of 4.9, 5.3, and 5.7 log10 CFU/mL (at 407, 632 and 668 kJ/L, respectively) and
a higher reduction of 7.1 log10 (at 815 kJ/mL) in milk with that of thermal pasteurization. However,
combined processing of MF followed by PEF resulted in lower reductions of 4.1 (at 407 and
632 kJ/L), 4.4 (at 668 kJ/L) and 4.8 (at 815 kJ/L) log10 CFU/mL in microbial population of the
milk microorganisms but was analogous to that of thermal pasteurization. Therefore, the results
indicate the importance of sequence of treatments in achieving higher inactivation efficiencies.
Both PEF/MF and thermally pasteurized milk expressed an overall shelf stability of 7 days based
on total aerobic counts. Thus, this combination has potential as an alternative treatment to thermal
pasteurization in milk provided with further extensive studies (Walkling-Ribeiro et al., 2011). The
combination of cross-flow microfiltration (CFMF) followed by PEF achieved similar antimicrobial
effect as that of CFMF followed by high-temperature short-time treatment (HTST). Also, PEF/
CFMF sequence resulted in higher inactivation of mesophilics compared to CFMF/PEF and HTST
indicating the vital role of order of treatments. The shelf life was comparable for CFMF/PEF and
HTST after 7 days (Gonzalez, 2010).
17.4.16 pulsed Electric Field and high hydrostatic pressure
The combination of two comparatively well-established non-thermal processing technologies,

HHP and PEF is a promising approach in the exploration for new alternatives for the preservation
of food products. Neither PEF nor HHP is able to achieve inactivation of spores alone. Therefore, to
disrupt the dormancy of the spores an intermediate step is reasonably required by which germination
of spores can be achieved for later potential HIPEF inactivation. In this context, HHP is a promising
technique which can germinate the spores at low doses without affecting the quality of food prod-
uct to be treated. Germinated spores become susceptible and might be inactivated by further mild
treatments such as PEF and heat. This hypothesis was confirmed by Pagan et al. (1998) in which
results revealed that no inactivation of Bacillus subtilis spores was achieved when only HIPEF was
used (60 kV/cm, 75 pulses). HHP (1500 atm, 30 min, and 40°C) resulted in the germination of more
than five log cycles of spores, making them sensitive to subsequent HIPEF at temperatures above
40°C. Thus, the combination is considered an attractive alternative to intensive thermal treatment
to inactivate spores. The sequential processing of PEF and HHP achieved a 7.1-log reduction in the
viable spore counts of Bacillus subtilis in the buffer solution (pH 3.3) and 6.5 log reduction in the
orange juice (pH 3.9), whereas, in contrast to previous finding, sequential processing of HHP and
PEF resulted in increase in viable spore counts after the HHP treatment due to the subsequent PEF
treatment which may be associated with the formation of H-spores (Sasagawa et al., 2006). In a
study, cut apples maintain the initial microbial population right after treatment (<1.0 × 101 CFU/mL)
for up to 2 months of storage at 5°C when treated with combinations of non-thermal hurdles such as
HHP, PEF, and/or 60°Brix sucrose osmotic solution containing ascorbic acid (200 ppm), citric acid
(200 ppm), sodium benzoate (200 ppm) and potassium sorbate (200 ppm) (Shayanfar et al., 2014).
17.4.17 high hydrostatic pressure and Ultraviolet Light assisted

titanium dioxide photocatalysis (tiO2-UV photocatalysis)
TiO2-assisted ultraviolet photocatalysis (TUVP) treatment is a well-established technique for

disinfection of water (Kim et al., 2013). In recent years, the use of TUVP has gained considerable
attention towards its use in food decontamination (Chai et al., 2014; Yoo et al., 2015).
When UV radiation at energies larger than the band-gap energy of the TiO2 crystal is absorbed
by the catalyst, an electron is promoted to the conduction band which results in formation of a pair of
a negatively-charged free electron and a positively-charged electron hole (Maness et al., 1999). The
electron-hole pair has strong reducing and oxidizing activities which reacts with water to produce
hydroxyl radicals (OH). The electron in the conduction band subsequently reacts with oxygen to
yield superoxide anions (O2−). During this reaction formation of other reactive oxygen species (ROS)
including hydrogen peroxide (H2O2) are also reported in some investigations (Maness et al., 1999).
These reactive species are responsible for redox reactions thus exhibit excellent microbial activity
by causing complete oxidation of organic compounds (Liou and Chang, 2012). TiO2 photocatalysis
have shown antibacterial effects against broad spectrum microorganisms. The possible mechanism
of action involves damage to the cell functions and structures, chemical modification or cleavage
of the DNA, disorder in cell permeability, damage to polyunsaturated phospholipids however, the
mechanisms leading to inactivation of microorganisms is not well understood.
Recently, combination of TUVP and HHP was assessed in the line of hurdle approach. Chai
et al. (2014) investigated combination of TiO2-UV photocatalysis and HHP to inactivate naturally
occurring Bacillus cereus in freshly squeezed Angelica keiskei juice. The combination of TUVP
and HHP achieved highest inactivation efficiency as compared to HHP and TUVP alone. HHP
was found to be highly efficient for inactivation of naturally occurring yeasts and moulds, coliform
bacteria, and Pseudomonas in Angelica keiskei juice, reducing their levels to below the detection
limit however, HHP reduced B. cereus to only 2.20 log CFU/mL due to the presence of B. cereus
endospores in squeezed A. keiskei juice that are resistant to HHP. The combination treatment
of TUVP and HHP reduced the levels of all forms of naturally occurring microflora including
B. cereus to below the detection limit, however injured yet surviving population of B. cereus,
germinate, and multiplied to a level of 2.02 log CFU/mL during storage. The results revealed a
possible potential of this combination in future prospects. Also, a combination of TUVP and HHP
reduced population of E. coli 0157:147 in orange juice by 4.7 log CFU/mL (below detection limit)
when TUVP treatment of oranges was followed by HHP treatment of juice, indicating a resultant
synergistic inactivation effect (Yoo et al., 2015). A sequential combination of TUVP and HHP
was found to be successful in accomplishing synergistic effect by inactivating internalized murine
norovirus to below detection limit (5.5 log10 reduction), which was not achievable by either of the
treatments alone (Kim et al., 2017). Combination of TiO2-UV photocatalysis and HHP to inactivate
bacterial pathogens and yeast in commercial apple juice expressed in synergistic effect. Listeria
monocytogenes and Staphylococcus aureus displayed complete inactivation, Escherichia coli
O157:H7 and Salmonella typhimurium, were reduced by 7.1 and 7.2 log CFU/mL, respectively,
Saccharomyces cerevisiae count was reduced by 6.2 log CFU/mL. The research findings suggest
potential opportunities for this synergistic disinfection strategy in the beverage industry provided
meticulous studies are required to discover inactivation effects on other microorganisms in different
category of food matrices and its effect on other quality parameters of tested food product (Shahbaz
et al., 2016).
17.4.18 Ozone and Ultrasonication
The potential deterioration or alteration in the sensory attributes in food products such as discol-
oration in food products due to surface oxidation and the adverse effects on nutrients (vitamins and
amino acids) owing to the highly reactive nature of ozone limits its applications in food products.
Therefore, coupling ozonation with other non-thermal technologies can overcome these limitations.
In a study conducted by Jyoti and Pandit (2004) on water disinfection by combination of ozone and
cavitation processing resulted in synergistic effect with reduction in required concentration of ozone
for disinfection to half or one-third, due to faster sedimentation of organic matter by ultrasound
resulting in reduced uptake of ozone. Also, the disinfection efficiency of hydrodynamic cavitation
was reported to enhance when used with ozone (Priyanka et al., 2014).
17.5 CUrrENt StatUS IN hUrDLE apprOaCh tOWarDS

NON-thErMaL prOCESSING tEChNOLOGIES at
LaBOratOrY aND COMMErCIaL LEVEL
The food industry has long been occupied in a hunt for a model non-thermal food processing
alternative to traditional thermal processes such as pasteurization and sterilization. However, none
of these technologies has yet proved to be a milestone technology as thermal processing. Consumer’s
preference in a processed food product has been shifted towards freshness and healthiness. To
achieve these objectives in addition to food safety, combination effects or hurdle approach is the
focus of many of the recent developments in non-thermal food processing technologies. Combining
non-thermal processing technologies with conventional preservation treatments has been widely
investigated in order to increase the lethal effect, while the information on combination effects of two
or more non-thermal technologies is scarce. In addition to positive effect of non-thermal technologies
in achieving food safety, these technologies are proved to be environmentally friendly due to the lack
of residual compounds and the absence of applied chemical disinfectants and preservatives. Based
on the available research findings, these technologies can be used as pre-treatments, in simultaneous
application with other technologies and in the final steps of minimal processing, depending on the
compatibility of different non-thermal technologies with respect to the food product to be treated.
These technologies may prove to be beneficial to food industry as energy-efficient and waste-free
technologies. Significant research has been carried out for estimating the potential of non-thermal
processing technologies; however, more research and development activities are required for better
understanding and optimization of processing parameters in order to enable the scale-up these pro-
cesses for food industry.
Non-thermal technology such as HHP and irradiation has already been applied to good number
of products at commercial level and is well established in the industry, other technologies such
as PEF and US are continue to be developed and evaluated for commercial application, while
technologies such as cold plasma, EBP, etc. are continue to be assessed at laboratory scale studies.
Rigorous research is required to understand the mechanism of action (multi-target) when these
technologies are applied in hurdle concept for optimizing the parameters of applied combinations
to specific food products at commercial level.
At present, HHP in combination with vacuum packaging is effectively utilized by the meat
industry in raw as well as ready-to-eat products, having similar appearance and characteristics of
regular products. The process application involves vacuum packaging followed by high-pressure
exposure to reduce the probability of post-processing or secondary contamination while

maintaining the freshness of the meat products with extended shelf life. Major market players of
meat and poultry manufacturers of the USA namely Hormel’s Natural Choice, Applegate Farms,
and Perdue Short Cuts have applied HHP technology to various meat products. Furthermore, HHP
technology effectively inhibits pathogens in ready-to-eat meat products and fresh meat products and
its legitimate use in eliminating Listeria monocytogenes in processed meats is approved by USDA
Food Safety and Inspection Service. In HHP-treated cured meat products, the reduced salinity
of the product and the elimination of need of addition of chemical antibacterial agents, results in
wholesome and natural like meat products that can successfully meet the necessities of Clean Label
enabling the widespread adoption of HHP technology among food industry to further make a way
into overseas markets (Huang et al., 2017).
The growth of another non-thermal technology i.e. irradiation has also witnessed significant hike
due to increased awareness and interest in the technology. United States witnessed marked increment
in annual sales (15 million–18 million pounds) of irradiated ground beef and poultry. Huisken Meat
Company, Schwan’s and Omaha Steaks are leading manufacturers of ground beef in the USA.
These highly respected food companies have made the decision to irradiate ground beef because of
serious concerns about food-borne illness from E. coli and Salmonella. Also, it was estimated that
in 2010 about 15,000 metric tons (35 million pounds) of irradiated fresh produce was consumed
in the United States and one-third of commercial spices (about 175 million pounds) are irradiated
and consumed in the United States (Eustice, 2011). Therefore, successful commercial application
of this technology suggests its utilization in hurdle approach to meet consumer’s expectations of
fresher and nutritious food products. Some of the possible non-thermal combinations of these
technologies are already discussed in this chapter and apart from these, research investigations
uttered productive results of irradiation in combination of packaging techniques such as vacuum
packaging and modified atmosphere packaging.
17.6 FUtUrE SCOpE IN hUrDLE apprOaCh tOWarDS

NON-thErMaL prOCESSING tEChNOLOGIES
Non-thermal processing technologies may serve as an alternative to thermal processing (pasteuri-

zation) when used in combination strategy, thus fulfilling the requirement of modern consumer’s by
providing safe, nutritious, shelf-stable, and organoleptically sound food products. Conversely, high
capital investment and limited throughput are major limitations in commercialization of most non-
thermal technologies. Therefore, with large-scale industrial adaptation, technology advancement, and
increasing consumer acceptance, it is projected that the cost of processing will definitely come down.
Despite the promising effects of some of the non-thermal technologies such as US, these technologies
are not followed to the net level of scaling-up at pilot scale. Consequently, further research is needed
to assess the combined effects of these technologies while targeting the scale-up level. Furthermore,
investigations focusing on nutritional and sensory parameters in addition to food safety are the need
of the hour. Moreover, many of the concerted efforts of researchers, are limited to assess the efficacy
of non-thermal technologies, on buffers or ideal solutions while, food matrices exert different level of
complications due to compositional interactions. Therefore, for real applications this component needs
to be considered for process optimization in meeting food safety.
Few non-thermal technologies such as high-pressure processing, PEF processing, irradiation,
US and ozonation have settled themselves in commercial application to a certain extent. However,
the concept of hurdle technology was not well addressed at commercial level, although they are
utilizing inherent barriers in achieving desired objectives for example in case of juices the inherent
low pH proves to be added advantage in attaining desired goals of food safety during processing
as well as storage. The universally accepted, optimum combinations of processing parameters
are still awaited as in case of thermal pasteurization and sterilization. Therefore, painstaking
research investigations are required in terms of technicality and biological effects of various
combinations to achieve this goal. Other non-thermal technologies are considered as emerging
technologies as these are in infancy stage in terms of commerciality and continued to be assessed
extensively at laboratory and pilot scale. Desired objective in hurdle concept towards non-thermal
processing technologies can only be accomplished by selecting the barriers based on technical
compatibility of selected treatments. Since, these technologies cannot be applied to all types of
food products. These technologies have limitations in application as these are very much dependent
on product characteristics. For example, pulsed light, UV, and ozonation are restricted to surface
decontamination of food products and, for sterilization of packaging material, PEF and US are
better suited to liquid food products, whereas HHP cannot be applied to porous food products.
Therefore, these factors play an important role in selection of barriers. On the other hand, economic
analysis of the processing treatment is also an imperative aspect especially in developing countries
while applying hurdle approach in non-thermal processing as most of the technologies need high
capital investment in creating the facility. Promising outcome of several research investigations
mentioned in this chapter indicates intelligent opportunities of combination technologies in food
industry, while antagonistic effect in few research investigations cannot be ignored and outcomes
can be considered in future selection of hurdles. Many of the combinations showed effective results
when non-thermal technologies were combined with heat; therefore, potential investigations require
painstaking efforts to make heat obsolete in realistic scenario. Consequently, the real potential of
non-thermal food processing technologies will not be realized unless advanced levels of research
and development programmes are initiated.
rEFErENCES
Abid, M., Jabbar, S., Hu, B., Hashim, M. M., Wua, T., Wu, Z., Khan, M. A., and Zeng, X. (2014). Synergistic
impact of sonication and high hydrostatic pressure on microbial and enzymatic inactivation of apple
juice. LWT – Food Science and Technology. 59(1): 70–76.
Aertsen, A., De Spiegeleer, P., Vanoirbeek, K., Lavilla, M., and Michiels, C. W. (2005). Induction of oxidative
stress by high hydrostatic pressure in Escherichia coli. Applied and Environmental Microbiology. 71:
2226–2231.
Ahmed, F. I. K., and Russell, C. (1975). Synergism between ultrasonic waves and hydrogen peroxide in the
killing of micro-organisms. Journal of Applied Bacteriology. 39: 31–40.
Ait-Ouazzou, A., Espina, L., García-Gonzalo, D., and Pagán, R. (2013). Synergistic combination of physical
treatments and carvacrol for Escherichia coli O157:H7 inactivation in apple, mango, orange, and tomato
juices. Food Control. 32: 159–167.
Alahakoon, A. U., Jayasena, D. D., Jung, S., Kim, S. H., Kim H. J., and Jo, C. (2015). Effects of electron beam
irradiation and high pressure treatment combined with citrus peel extract on seasoned chicken breast
meat. Journal of Food Processing and Preservation. 39: 2332–2339.
Alpas, H., Kalchayanand, N., Bozozlu, F., and Ray, B. (2000). Interactions of high hydrostatic pressure, pres-
surization temperature and pH on death and injury of pressure-resistant and pressure sensitive strains of
foodborne pathogens. International Journal of Food Microbiology. 60: 33–42.
Alpas, H., Kalchayanand, N., Bozoglu, F., Sikes, A., Dunne, C. P., and Ray, B. (1999). Variation in resistance to
hydrostatic pressure among strains of food-borne pathogens. Applied and Environmental Microbiology.
65: 4248–4251.
Amanatidou, A., Schlüter, O., Lemkau, K., Gorris, L. G. M., Smid, E. J., and Knorr, D. (2000). Effect of com-
bined application of high pressure treatment and modified atmosphere on the shelf life of fresh Atlantic
salmon. Innovative Food Science and Emerging Technologies. 1: 87–98.
Amiali, M., and Ngadi, M. O. (2012). Microbial decontamination of food by pulsed electric fields (PEFs). In:
Microbial Decontamination in the Food Industry, Novel Methods and Applications, pp. 407–449. A
volume in Woodhead Publishing Series in Food Science, Technology and Nutrition.
Anaya-Esparza, L. M., Velázquez-Estrada, R. M., Roig, A. X., García, H. S., Sayago-Ayerdi, S. G., and
Montalvo-González, E. (2017). Thermosonication: An alternative processing for fruit and vegetable
juices. Trends in Food Science and Technology. 61: 26–37.
Awad, T. S., Moharram, H. A., Shaltout, O. E., Asker, D., and Youssef, M. M. (2012). Applications of ultra-
sound in analysis, processing and quality control of food: A review. Food Research International. 48:
410–427.
Balasubramanian, S., and Balasubramaniam, V. M. (2003). Compression heating influence of pressure-
transmitting fluids on bacteria inactivation during high pressure processing. Food Research
International. 36(7): 661–668.
Balasubramaniam, V. M., Balasubramaniam, S., and Reddy, N. R. (2001). Effect of pH and temperature on
spore inactivation during high pressure processing. IFT Annual Meeting Book of Abstracts, Session
59H-5.
Bazhal, M. I., Ngadi, M. O., and Raghavan, V. G. S. (2006). Kinetics of Escherichia coli in liquid whole egg
using combined PEF and thermal treatments. LWT – Food Science and Technology. 39(4): 420–426.
Bermúdez-Aguirre, D., Corradini, M. G., Mawson, R., and Barbosa-Cánovas, G. V. (2009). Modeling the
inactivation of Listeria innocua in raw whole milk treated under thermo-sonication. Innovative Food
Science and Emerging Technologies. 10: 172–178.
Bermudez-Aguirre, D., Dunne, C. P., and Barbosa-Canovas, G. V. (2012). Effect of processing parameters on
inactivation of Bacillus cereus spores in milk using pulsed electric fields. International Dairy Journal.
24(1): 13–21.
Birmpa, A., Sfika, V., and Vantarakis, A. (2013). Ultraviolet light and ultrasound as non-thermal treatments
for the inactivation of microorganisms in fresh ready-to-eat foods. International Journal of Food
Butz, P., Funtenberger, S., Haberditzl, T., and Tauscher, B. (1995). High pressure inactivation of
Byssochlamis nivea ascospores and other heat-resistant moulds. LWT – Food Science and Technology.
29: 404–410.
Butz, P., and Tauscher, B. (2002). Emerging technologies: Chemical aspects. Food Research International.
35(2/3): 279–284.
Calderon-Miranda, M. L., Barbosa-Canovas, G. V., and Swanson, B. G. (1999). Inactivation of Listeria
innocua in skim milk by pulsed electric fields and nisin. International Journal of Food Microbiology.
51: 19–30.
Caminiti, A. M., Noci, F., Munoz, A., Whyte, P., Morgan, D. J., Cronin, D. A., and Lyng, J. G. (2011). Impact of
selected combinations of non-thermal processing technologies on the quality of an apple and cranberry
juice blend. Food Chemistry. 124: 1387–1392.
Caminiti, I. M., Noci, F., Morgan, D. J., Cronin, D. A., and Lyng, J. G. (2012). The effect of pulsed elec-
tric fields, ultraviolet light or high intensity light pulses in combination with manothermosonication
on selected physico-chemical and sensory attributes of an orange and carrot juice blend. Food and
Bioproducts Processing. 90: 442–448.
Cappelletti, M., Ferrentino, G., and Spilimbergo, S. (2014). Supercritical carbon dioxide combined with high
power ultrasound: An effective method for the pasteurization of coconut water. Journal of Supercritical
Fluids. 92: 257–263.
Carlez, A., Rosec, J. P., Richard, N., and Cheftel, J. (1993). High pressure inactivation of Citrobacter freundii,
Pseudomonas fluorescens, and Listeria innocua in inoculated minced beef. LWT – Food Science and
Chai, C., Lee, J., Lee, Y., Na, S., and Park, J. (2014). A combination of TiO2-UV photocatalysis and high
hydrostatic pressure to inactivate Bacillus cereus in freshly squeezed Angelica keiskei juice. LWT –
Food Science and Technology. 55(1): 104–109.
Char, C. D., Mitilinaki, E., Guerrero, S. R., and Alzamora, S. M. (2010). Use of high-intensity ultrasound and
UV-C light to inactivate some microorganisms in fruit juices. Food Bioprocess Technology. 3: 797–803.
Chun, H. H., and Song, K. B. (2014). Optimisation of the combined treatments of aqueous chlorine dioxide,
fumaric acid and ultraviolet-C for improving the microbial quality and maintaining sensory quality of
common buckwheat sprout. International Journal of Food Science and Technology. 49(1): 121–127.
Ciccolini, L., Taillandier, P., Wilhem, A. M., Delmas, H., and Strehaiano, P. (1997). Low frequency thermo-
ultrasonication of Saccharomyces cerevisiae suspensions: Effect of temperature and of ultrasonic
power. Chemical Engineering Journal. 65: 145–149.
Condon, S., Álvarez, I., and Gayán, E. (2014). Pulsed UV light. In: Encyclopedia of Food Microbiology,
pp. 974–981. Batt, C. A., and Tortorello, M. L. Eds. 2nd ed. Amsterdam, the Netherlands: Elsevier.
Coronel, C. P., Jiméneza, M. T., López-Malo, A., and Palou, E. (2011). Modelling thermosonication inactiva-
tion of Aspergillus flavus combining natural antimicrobial at different pH. Procedia – Food Science. 1:
1007–1014.
Cregenzán-Alberti, O., Halpin, R. M., Whyte, P., Lyng, J. G., and Noci, F. (2015). Study of the suitability
of the central composite design to predict the inactivation kinetics by pulsed electric fields (PEF) in
Escherichia coli, Staphylococcus aureus and Pseudomonas fluorescens in milk. Food and Bioproducts
Processing. 95: 313–322.
Crawford, Y. J., Murano, E. A., Olson, D. G., and Shenoy, K. (1996). Use of high hydrostatic pressure and
irradiation to eliminate Clostridium sporogenes spores in chicken breast. Journal of Food Protection.
59: 711–715.
Cullen, P. J., Tiwari, B. K., and Valdramidis, V. P. (2012). Novel Thermal and Non-Thermal Technologies for
Fluid Foods. Academic Press, Oxford, UK.
Dutreux, N., Notermans, S., Wijtzes, T., Góngora-Nieto, M. M., Barbosa-Cánovas, G. V., and Swanson, B.
G. (2000). Pulsed electric fields inactivation of attached and free-living Escherichia coli and Listeria
innocua under several conditions. International Journal of Food Microbiology. 54: 91–98.
Duong, T., and Balaban, M. (2014). Optimisation of the process parameters of combined high hydrostatic
pressure and dense phase carbon dioxide on enzyme inactivation in feijoa (Acca sellowiana) puree
using response surface methodology. Innovative Food Science and Emerging Technologies. 26:
93–101.
Earnshaw, R. G., Appleyard, J., and Hurst, R. M. (1995). Understanding physical inactivation processes:
Combined preservation opportunities using heat, ultrasound and pressure. International Journal of
El’piner, E. A. (1964). Ultrasounds: Physical, Chemical and Biological Effects. New York: Consultants
Bureau.
Eshtiaghi, M. N., and Knorr, D. (1993). Potato cubes response to water blanching and high hydrostatic pres-
sure. Journal of Food Science. 58(6): 1371–1374.
Espina, L., García-Gonzalo, D., Laglaoui, A., Mackey, B. M., and Pagán, R. (2013). Synergistic combinations
of high hydrostatic pressure and essential oils or their constituents and their use in preservation of fruit
juices. International Journal of Food Microbiology. 161: 23–30.
Eustice, R. (2011). Food irradiation: A global perspective & future prospects. http://ansnuclearcafe.
org/2011/06/09/food-irradiation-a-global-perspective-future prospects/#sthash.imZeMV3N.dpuf.
Evelyn, and Silva, F. V. M. (2015). Use of power ultrasound to enhance the thermal inactivation of Clostridium
perfringens spores in beef slurry. International Journal of Food Microbiology. 206: 17–23.
Evelyn, and Silva, F. V. M. (2016). High pressure processing pretreatment enhanced the thermosonication
inactivation of Alicyclobacillus acidoterrestris spores in orange juice. Food Control. 62: 365–372.
Ferrario, M., Alzamora, S. M., and Guerrero, S. (2015). Study of the inactivation of spoilage microorganisms
in apple juice by pulsed light and ultrasound. Food Microbiology. 46: 635–642.
FDA. (1996). Code of Federal Regulations. 21CFR179.41.
Flores-Cervantes, D. X., Palou, E., and Lopez-Malo, A. (2013). Efficacy of individual and combined UVC
light and food antimicrobial treatments to inactivate Aspergillus flavus or A. niger spores in peach
nectar. Innovative Food Science & Emerging Technologies. 20: 244–252.
Francesco, D., Enrico, M., Paola, M., Gianpiero, P., Dang, V. K., Stephane, S., Monique, L., and Giovanna,
F. (2015). Green beans preservation by combination of a modified chitosan based-coating containing
nanoemulsion of mandarin essential oil with high pressure or pulsed light processing. Postharvest
Biology and Technology. 106: 21–32.
Fritsch, J., and Moraru, C. I. (2008). Development and optimization of a carbon dioxide-aided cold
microfiltration process for the physical removal of microorganisms and somatic cells from skim milk.
Journal of Dairy Science. 91: 3744–3760.
García, D., Gómez, N., Mañas, P., Condón, S., Raso, J., and Pagán, R. (2005). Occurrence of sublethal injury
after pulsed electric fields depending on the micro-organism, the treatment medium pH and the intensity
of the treatment investigated. Journal of Applied Microbiology. 99(1): 94–104.
Garcia, M. L., Burgos, J., Sanz, B., and Ordonez, J. (1989). Effect of heat and ultrasonic waves on the survival
of two strains of Bacillus subtilis. Journal of Applied Microbiology. 67: 619–628.
García-García, R., Escobedo-Avellaneda, Z., Tejada-Ortigoza, V., Martín-Belloso, O., Valdez-Fragoso,

A., and Welti-Chanes, J. (2015). Hurdle technology applied to prickly pear beverages for inhibiting
Saccharomyces cerevisiae and Escherichia coli. Letters in Applied Microbiology. 60: 558–564.
Garcia-Gonzalez, L., Geeraerd, A. H., Spilimbergo, S., Elst, K., Van Ginneken, L., Debevere, J., Van Impe,
J. F., and Devlieghere, F. (2007). High pressure carbon dioxide inactivation of microorganisms in foods:
The past, the present and the future. International Journal Food Microbiology. 117: 1–28.
Garcia-Graells, C., Hauben, E. J. A., and Michiels, C. W. (1998). High-pressure inactivation and sublethal
injury of pressure-resistant Escherichia coli mutants in fruit juices. Applied and Environmental
Garcia-Graells, C., Masschalck, B., and Michiels, C. W. (1999). Inactivation of Escherichi coli in milk by
high-hydrostatic-pressure treatment in combination with antimicrobial peptides. Journal of Food
Protection. 62: 1248–1254.
García-Parra, J., González-Cebrino, F., Delgado, J., Cava, R., and Ramírez, R. (2016). High pressure assisted
thermal processing of pumpkin purée: Effect on microbial counts, color, bioactive compounds and
polyphenoloxidase enzyme. Food and Bioproducts Processing. 98: 124–132.
Gayán, E., García-Gonzalo, D., Álvarez, I., and Condón, S. (2014). Resistance of Staphylococcus aureus to
UV-C light and combined UV-heat treatments at mild temperatures. International Journal of Food
Gayán, E., Serrano, M. J., Monfort, S., Álvarez, I., and Condón, S. (2012). Combining ultraviolet light and
mild temperatures for the inactivation of Escherichia coli in orange juice. Journal of Food Engineering.
113: 598–605.
Giese, N., and Darby, J. (2000). Sensitivity of microorganisms to different wavelengths of UV light:
Implications on modeling of medium pressure UV systems. Water Research. 34: 4007–4013.
Giner, J., Gimeno, V., Espachs, A., Elez, P., Barbosa-Cánovas, G. V., and Martín, O. (2000). Inactivation of
tomato (Licopersicon esculentum Mill.) pectin methylesterase by pulsed electric fields. Innovative Food
Science and Emerging Technologies. 1: 57–69.
Giovanna, F., and Sara, S. (2015). High pressure carbon dioxide combined with high power ultrasound
pasteurization of fresh cut carrot. Journal of Supercritical Fluids. 105: 170–178.
Gómez-López, V. M., Ragaert, P., Debevere, J., and Devlieghere, F. (2007). Pulsed light for food
decontamination: A review. Trends in Food Science and Technology. 18(9): 464–473.
Gonzalez, O. R. (2010). Hurdle technologies: Microbial inactivation by pulsed electric fields during milk
processing. A Thesis Presented to the Faculty of Graduate Studies of the University of Guelph.
Gosch, T., Apprich, S., Kneifel, W., and Novalin, S. (2014). A combination of microfiltration and high pres-
sure treatment for the elimination of bacteria in bovine colostrums. International Dairy Journal. 34:
41–46.
Gould, G. W. (1996). Industry perspectives on the use of natural antimicrobials and inhibitors for food
applications. Journal of Food protection. 59(3): 82–86.
Gould, G. W., and Sale, J. H. (1970). Initiation of germination of bacterial spores by hydrostatic pressure.
Journal of General Microbiology. 60: 335–346.
Gouma, M., Gayan, E., Raso, J., Condon, S., and Alvarez, I. (2015). Inactivation of spoilage yeasts in apple
juice by UV–C light and in combination with mild heat. Innovative Food Science and Emerging
Technologies. 32: 146–155.
Grahl, T., and Markl, H. (1996). Killing of microorganisms by pulsed electric fields. Applied Microbiology
and Biotechnology. 45: 148–157.
Guerrero, S., Lo´pez-Malo, A., and Alzamora, S. M. (2001). Effect of ultrasound on the survival of
Saccharomyces cerevisiae: Influence of temperature, pH and amplitude. Innovative Food Science and
Emerging Technologies. 2: 31–41.
Guerrero, S., Tognon, M., and Alzamora, S. M. (2005). Response of Saccharomyces cerevisiae to the combined
action of ultrasound and low weight chitosan. Food Control. 16: 131–139.
Guzel-Seydim, Z. B., Bever, P. I. J., and Greene, A. K. (2004b). Efficacy of ozone to reduce bacterial
populations in the presence of food components. Food Microbiology. 21: 475–479.
Guzel-Seydim, Z. B., Greene, A. K., and Seydim, A. C. (2004a). Use of ozone in the food industry. LWT –
Food Science and Technology. 37(4): 453–460.
Ha, J. H., Kim, H. J., and Ha, S. D. (2012). Effect of combined radiation and NaOCl/ultrasonication on
reduction of Bacillus cereus spores in rice. Radiation Physics and Chemistry. 81: 1177–1180.
Hagenmaier, R. D., and Baker, R. A. (1998). Microbial population of shredded carrot in modified atmosphere
packaging as related to irradiation treatment. Journal of Food science. 68: 162–164.
Halander, I. M., Wright, A. V., and Mattila-Sandholm, T. M. (1997). Potential of lactic acid bacteria and novel
antimicrobials against Gram-negative bacteria. Trends in Food Science and Technology. 8: 151–156.
Halim, H., Noranizan, M., Sobhi, B., Sew, C. C., Karim, R., and Osman, A. (2012). Nonthermal pasteurization
of pitaya (Hylocereus polyrhizus) juice using the hurdle concept. International Journal of Food research.
19: 1457–1461.
Hauben, K. J. A., Wuytack, E. Y., Soontjens, C. C. F., and Michiels, C. W. (1996). High-pressure transient
sensitization of Escherichia coli to lysozyme and nisin by disruption of outer-membrane permeability.
Journal of Food protection. 59: 350–355.
Hayakawa, I., Kanno, T., Tomita, M., and Fujio, Y. (1994). Application of high pressure for spore inactivation
and protein denaturation. Journal of Food Science. 59: 159–163.
Heinz, V., Toepfl, S., and Knorr, D. (2003). Impact of temperature on lethality and energy efficiency of
apple juice pasteurization by pulsed electric fields treatment. Innovative Food Science and Emerging
Technologies. 4(2): 167–175.
Hernandez, A., and Cano, M. P. (1998). High-pressure and temperature effects on enzyme inactivation in
tomato puree. Journal of Agriculture and Food Chemistry. 46: 266–270.
Hendrickx, M., Ludikhuyze, L., Van den Broeck, I., and Weemaes, C. (1998). Effects of high pressure on
enzymes related to food quality. Trends in Food Science and Technology. 9: 197–203.
Ho, S. H., Mittal, G. S., and Cross, J. D. (1997). Effects of high electric pulses on the activity of selected
enzymes. Journal of Food Engineering. 31: 69–84.
Ho, S. Y., and Mittal, G. S. (2000). High voltage pulsed electrical field for liquid food pasteurization. Food
Review International. 16: 395–434.
Hoover, D. G., Metrick, C., Papineau, A. M., Farkas, D. F., and Knorr, D. (1989). Biological effects of high
hydrostatic pressure on food microorganisms. Food Technology. 43: 99–107.
Huang, H. W., Wu, S. J., Lu, J. K., Shyu, Y. T., and Wang, C. Y. (2017). Current status and future trends of
high-pressure processing in food industry. Food Control. 72: 1–8.
Izabela, P., Barbara, S., Sylwia, S., Sylwester, J. R. (2017). Treatment with high hydrostatic pressure and
supercritical carbon dioxide to control Alicyclobacillus acidoterrestris spores in apple juice. Food
Control. 73: 24–30.
Jaeger, H., Meneses, N., and Knorr, D. (2014). Food technologies: Pulsed electric field technology. In
Encyclopedia of Food Safety, pp. 239–244. Y. Motarjemi ed. 3rd Edn., London: Academic Press.
Jin, T., Zhang, H., Hermawan, N., and Dantzer, W. (2009). Effects of pH and temperature on inactivation of
Salmonella typhimurium DT104 in liquid whole egg by pulsed electric fields. International Journal of
Food Science and Technology. 44: 367–372.
Jin, Z. T., Su, Y., Tuhela, L., Singh, B., and Zhang, Q. H. (1998). Inactivation of Bacillus subtilis using high
voltage pulsed electric fields and ultrasonication. IFT Annual Meeting Book of Abstracts, 1998, Session
59C-15.
Juven, B., Kanner, J., Schved, F., and Weisslowicz, H. (1994). Factors that interact with the antibacterial action
of thyme essential oil and its active constituents. Journal of Applied Bacteriology. 76: 626–631.
Jyoti, K. K., and Pandit, A. B. (2004). Ozone and cavitation for water disinfection. Biochemical Engineering
Journal. 18: 9–19.
Kalchayanand, N., Sikes, A., Dunne, P., and Ray, B. (1998a). Factors influencing death and injury of food
borne pathogens by hydrostatic pressure-pasteurization. Food Microbiology. 15: 207–214.
Kalchayanand, N., Sikes, A., Dunne, C. P., and Ray, B. (1998b). Interaction of hydrostatic pressure, time and
temperature of pressurization and pediocin AcH on inactivation of foodborne bacteria. Journal of food
protection. 61: 425–431.
Khadre, M. A., Yousef, A. E., and Kim, J. G., (2001). Microbiological aspects of ozone applications in food:
A review. Journal of Food Science. 65: 521–528.
Kim, A. Y., and Thayer, D. W. (1996). Mechanism by which gamma irradiation increases the sensitiv-
ity of Salmonella typhimurium ATCC 14028 to heat. Applied Environmental Microbiology. 62:
1759–1763.
Kim, H. J., Mingu, K., and Cheorun, J. (2012). Combined effects of electron beam irradiation and addition
of onion peel extracts and flavoring on microbial and sensorial quality of pork jerky. CNU Journal of
Agricultural Science. 39(3): 341–347.
Kim, H. J., Mingu, K., Yong, H. I., Bae, Y. S., Jung, S., and Jo, C. (2013). Synergistic effects of electron-
beam irradiation and leek extract on the quality of pork jerky during ambient storage. Asian-Australian
Journal of Animal Science. 26(4): 596–602.
Kim, J. G., Yousef, A. E., and Khadre, M. A. (2003). Ozone and its current and future application in the food
industry. Advances in Food and Nutrition Research. 45: 167–218.
Kim, S., Ghafoor, K., Lee, J., Feng, M., Hong, J., Lee, D., and Park, J. (2013). Bacterial inactivation in
water, DNA strand breaking, and membrane damage induced by ultraviolet assisted titanium dioxide
photocatalysis. Water Research. 47(13): 4403–4411.
Kim, S. H., Shahbaz, H. M., Park, D., Chun, S., Lee, W., Oh, J. W., Lee, D. U., and Park, J. (2017). A combined
treatment of UV-assisted TiO2 photocatalysis and high hydrostatic pressure to inactivate internalized
murine norovirus. Innovative Food Science and Emerging Technologies. 39: 188–196.
Koutchma, T. (2009). Advances in ultraviolet light technology for non-thermal processing of liquid foods.
Food Bioprocess Technology. 2: 138–155.
Knorr, D., Froehling, A., Jaeger, H., Reineke, K., Schlueter, O., and Schoessler, K. (2011). Emerging
technologies in food processing. Annual Review of Food Science and Technology. 2: 203–235.
Kumar, G. D., Williams, R. C., Sumner, S. S., and Eifert, J. D. (2016). Effect of ozone and ultraviolet light on
Listeria monocytogenes populations in fresh and spent chill brines. Food Control. 59: 172–177.
Labbé, R. G., Juneja, V. K., and Blascheck, H. P. (2014). Clostridium perfringens. In: Encyclopedia of
Food Microbiology, pp. 433–445. Batt, C. A., and Tortorello, M. L., Eds., 2nd edn. Amsterdam, the
Netherland: Elsevier.
Lee, H., Zhou, B., Liang, W., Feng, H., and Martin, S. E. (2009). Inactivation of Escherichia coli cells with
sonication, manosonication, thermosonication, and manothermosonication: Microbial responses and
kinetics modeling. Journal of Food Engineering. 93: 354–364.
Lee, M., Sebranek, J. G., Olson, D. G., and Dickson, J. S. (1996). Irradiation and packaging of fresh meat and
poultry. Journal of Food Protection. 59: 62–72.
Leistner, L. (1992). Food preservation by combined methods. Food Research International. 25: 151–158.
Leistner, L. (1995). Principles and applications of hurdle technology. In: New Methods of Food Preservation,
pp. 1–21. G. W. Gould., Eds., London, UK: Blackie Academic & Professional.
Leistner, L. (1999). Combined methods for food preservation. In: Handbook of Food Preservation, pp.
457 485. Rahman, M. S., Eds., New York: Marcel Dekker.
Leistner, L. (2000). Basic aspects of food preservation by hurdle technology. International Journal of Food
Leistner, L., and Gould, G. W. (2002). Hurdle Technologies: Combination Treatments for Food Stability,
Safety and Quality. New York: Kluwer Academic/Plenum Publishers.
Linton, M., McClements, J. M., and Patterson, M. F. (1999). Survival of Escherichia coli O157: H7 during
storage in pressure-treated orange juice. Journal of Food Protection. 62(9): 1038–1040.
Liou, J., and Chang, H. (2012). Bactericidal effects and mechanisms of visible light-responsive titanium
dioxide photocatalysts on pathogenic bacteria. Archivum Immunologiae Et Therapiae Experimentalis
(Warsz). 60(4): 267–275.
Liu, X., Yousef, A. E., and Chism, G. W. (1997). Inactivation of Escherichia coli O157: H7 by the combination
of organic acids and pulsed electric field. Journal of Food Safety. 16: 287–299.
Lopez-Caballero, M. E., Pe´rez-Mateos, M., Borderias, J. A., and Montero, P. (2000). Extension of the shelf
life of prawns (Penaeus japonicus) by vacuum packaging and high-pressure treatment. Journal of Food
Protection. 63: 1381–1388.
Lopez, P., Sala, F. J., de la Fuente, J. L., Condon, S., Raso, J., and Burgos, J. (1994). Inactivation of peroxidase,
lipoxygenase, and polyphenol oxidase by manothermosonication. Journal of Agriculture and Food
Chemistry. 42: 252–256.
Luz, A. M., and Martínez de, M. I. (2015). Inactivation of Bacillus subtilis spores by combined pulsed light
and thermal treatments. International Journal of Food Microbiology. 214: 31–37.
Mackey, B. M., and Mãnas, P. (2008). Inactivation of Escherichia coli by high pressure. In: High Pressure
Microbiology, pp. 53–86. Michiels, C., Barlett, D. H., and Aertsen, A., Eds., Washington, DC: ASM
Press.
Maness, P. C., Smolinski, S., Blake, D. M., Huang, Z., Wolfrum, E. J., and Jacoby, W. A. (1999). Bactericidal
activity of photocatalytic TiO2 reaction: Toward an understanding of its killing mechanism. Applied and
Environmental Microbiology. 65(9): 4094–4098.
Markland, S. M., Kniel, K. E., Setlow, P., and Hoover, D. G. (2013). Nonthermal inactivation of heteroge-
neous and superdormant spore populations of Bacillus cereus using ozone and high pressure processing.
Innovative Food Science and Emerging Technologies. 19: 44–49.
Marquenie, D., Geeraerd, A. H., Lammertyn, J., Soontjens, C., Van Impe, J. F., and Michiels, C. W. (2003).
Combinations of pulsed white light and UV-C or mild heat treatment to inactivate conidia of Botrytis
cinerea and Monilia fructigena. International Journal of Food Microbiology. 85: 185–196.
Marquenie, D., Lammertyn, J., Geeraerd, A. H., Soontjens, C., Van Impe, J. F., Nicolaı̈, B. M., and Michiels,
C. W. (2002). Inactivation of conidia of Botrytis cinerea and Monilinia fructigena using UVC and heat
treatment. International Journal of Food Microbiology. 74 (1–2): 27–35.
Marquez, V. O., Mittal, G. S., and Griffiths, M. W. (1997). Destruction and inhibition of bacterial spores by
high voltage pulsed electric field. Journal of Food Science. 62: 399–401.
Masschalck, B., Garcia-Graells, C., Van Haver, E., and Michiels, C. W. (2000). Inactivation of high pressure
resistant Escherichia coli by lysozyme and nisin under high pressure. Innovative Food Science and
Emerging Technologies. 1: 39–49.
Mertens, B., and Deplace, G. (1993). Engineering aspects of high-pressure technology in the food industry.
Food Technology. 47: 164–169.
McNamee, C., Noci, F., Cronin, D. A., Lyng, J. G., Morgan, D. J., and Scannell, A. G. M. (2010). PEF based
hurdle strategy to control Pichia fermentans, Listeria innocua and Escherichia coli k12 in orange juice.
International Journal of Food Microbiology. 138: 13–18.
Millan-Sango, D., Garroni, E., Farrugia, C., Van Impe, J. F. M., and Valdramidis, V. P. (2016). Determination
of the efficacy of ultrasound combined with essential oils on the decontamination of Salmonella
inoculated lettuce leaves. LWT – Food Science and Technology. 73: 80–87.
Milani, E. A., and Silva, F. V. M. (2017). Ultrasound assisted thermal pasteurization of beers with different alcohol
levels inactivation of Saccharomyces cerevisiae ascospores. Journal of Food Engineering. 198: 45–53.
Miller, R. B. (2005). Electronic Irradiation of Foods. New York, Springer.
Mills, G., Earnshaw, R., and Patterson, M. F. (1998). Effects of high hydrostatic pressure on Clostridium
sporogenes spores. Letters in Applied Microbiology. 26: 227–230.
Montiel, R., Bravo, D., de Alba, M., Gaya, P., and Medina, M. (2012). Combined effect of high pressure
treatments and the lactoperoxidase system on the inactivation of Listeria monocytogenes in cold-
smoked salmon. Innovative Food Science and Emerging Technologies. 16: 26–32.
Moosekian, S. R., Jeong, S., Marks, B. P., and Ryser, E. T. (2012). X-Ray irradiation as a microbial intervention
strategy for food. Annual Review of Food Science and Technology. 3: 493–510.
Morris, C., Brody, A. L., and Wicker, L. (2007). Non-thermal food processing/preservation technologies: A
review with packaging implications. Packaging Technology and Science. 20: 275–286.
Mosqueda-Melgar, J., Raybaudi-Massilia, R. M., and Martín-Belloso, O. (2008). Combination of high intensity
pulsed electric fields with natural antimicrobials to inactivate pathogenic microorganisms and extend
the shelf-life of melon and watermelon juices. Food Microbiology. 25: 479–491.
Mossel, D. A. A. (1983). Essentials and perspectives of the microbial ecology of foods. In: Food Microbiology:
Advances and Prospects, pp. 1–45. Roberts, T. A., and Skinner, E. A. Eds., London, UK: Academic Press.
Muñoz, A., Palgan, I., Noci, F., Morgan, D. J., Cronin, D. A., Whyte, P., and Lyng, J. G. (2011). Combinations
of high intensity light pulses and thermosonication for the inactivation of Escherichia coli in orange
juice. Food Microbiology. 28: 1200–1204.
Murano, P. S., Murano, E. A., and Olson, D. G. (1998). Irradiated ground beef: Sensory and quality changes
during storage under various packaging conditions. Journal of Food Science. 63: 548–551.
Naidu, A. S. (2000). Lactoperoxidase. In: Natural Food Antimicrobial Systems, pp. 103–132. Naidu, A. S.,
Ed., Boca Raton, FL: CRC Press.
Ngadi, M., Jun, X., Smith, J., and Raghavan, G. S. V. (2004). Inactivation of Escherichia coli 0157: H7 in
poultry chiller water using combined ultraviolet light, pulsed electric field and ozone treatments.
International Journal of Poultry Science. 3(11): 733–737.
Ngadi, M. O., Yu, L., Amiali, M., and Ortega-Rivas, E. (2009). Food quality and safety issues during pulsed
electric field processing. In: Processing Effects on Safety and Quality of Foods, pp. 446–472. Ortega-
Rivas, E., Ed. Boca Raton, FL: CRC Press.
Nguyen, P., and Mittal, G. S. (2007). Inactivation of naturally occurring microorganisms in tomato juice using
pulsed electric field (PEF) with and without antimicrobials. Chemical Engineering and Processing:
Process Intensification. 46(4): 360–365.
Noci, F., Riener, J., Walkling, M., Cronin, D. A., Morgan, D. J., and Lying, J. G. (2008). Ultraviolet irradiation
and pulsed electric fields (PEF) in a hurdle strategy for the preservation of fresh apple juice. Journal of
Food Science. 85: 141–146.
Noci, F., Walkling-Ribeiro, M., Cronin, D. A., Morgan, D. J., and Lyng, J. G. (2009). Effect of thermosonication,
pulsed electric field and their combination on inactivation of Listeria innocua in milk. International
Dairy Journal. 19: 30–35.
Ogawa, H., Fukuhisa, K., Kubo, Y., and Fukumoto, H. (1990). Pressure inactivation of yeast, molds, and
pectinesterase in satsuma mandarin juice: Effects of juice concentration, pH, and organic acids, and
comparison with heat sanitation. Agricultural and Biological Chemistry. 54: 1219–1225.
Ordonez, J. A., Aguilera, M. A., Garcia, M. L., and Sanz, B. (1987). Effect of combined ultrasonic and heat
treatment (thermoultrasonication) on the survival of a strain of Staphylococcus aureus. Journal of Dairy
Science. 54: 61–67.
Pagan, R., Esplugas, S., Gongora-Nieto, M. M., Barbosa-Canovas, G. V., and Swanson, B. G. (1998).
Inactivation of Bacillus subtilis spores using high intensity pulsed electric fields in combination with
other food conservation technologies. Food Science and Technology International. 4: 33–44.
Pagan, R., Manas, P., Alvarez, I., and Condon, S. (1999). Resistance of Listeria monocytogenes to ultrasonic
waves under pressure at sublethal (manosonication) and lethal (manothermosonication) temperatures.
Palacios, P., Burgos, J., Hoz, L., Sanz, B., and Ordóñez, J. A. (1991). Study of substances released by
ultrasonic treatment from Bacillus stearothermophilus spores. Journal of Applied Bacteriology. 71:
445–451.
Palgan, I., Caminiti, I. M., Munoz, A., Noci, F., Whyte, P., Morgan, D. J., Cronin, D. A., and Lyng, J. G. (2011).
Combined effect of selected non-thermal technologies on Escherichia coli and Pichia fermentans
inactivation in an apple and cranberry juice blend and on product. International Journal of Food
Food Processing, pp. 279–306. Sun, D. W., Ed., London, UK: Academic Press.
Pandya, Y., Jewett, F., and Hoover, D. G. (1995). Concurrent effects of high hydrostatic pressure, acidity and
heat on the destruction and injury of yeasts. Journal of Food Protection. 58: 301–304.
Park, S. Y., Mizan, M. F. R., and Ha, S. D. (2016). Inactivation of Cronobacter sakazakii in head lettuce by
using a combination of ultrasound and sodium hypochlorite. Food Control. 60: 582–587.
Patterson, M. F., Quinn, M., Simpson, R., and Gilmour, A. (1995). Sensitivity of vegetative pathogens to high
hydrostatic pressure treatment in phosphate-buffered saline and foods. Journal of Food Protection. 58:
524–529.
Paul, P., Chawla, S. P., Thomas, P., Kesavan, P. C., Fotedar, R., and Arya, R. N. (1997). Effect of high
hydrostatic pressure, gamma-irradiation and combination treatments on the microbiological quality of
lamb meat during chilled storage. Journal of Food Safety. 16: 263–271.
Ponce, E., Pla, R., Sendra, E., Guamis, B., and Mor-Mur, M. (1998). Combined effect of nisin and high
hydrostatic pressure on destruction of Listeria innocua and Escherichia coli in liquid whole egg.
International Journal of Food Microbiology. 43: 15–19.
Prakash, A., Guner, A. R., Caporaso, F., and Foley, D. M. (2000). Effects of low-dose gamma irradiation on
the shelf-life and quality characteristics of cut romaine lettuce packaged under modified atmosphere.
Journal of Food Science. 65: 549–553.
Priyanka, B. S., Rastogi, N. K., and Tiwari, B. K. (2014). Opportunities and challenges in the application of
ozone in food processing. In: Emerging Technologies for Food Processing, pp. 335–358. Sun, D. W.,
Ed., 2nd edn. London, UK: Elsevier, Academic Press.
Proulx, J., Sullivan, G., Marostegan, L. F., VanWees, S., Hsu, L. C., and Moraru, C. I. (2017). Pulsed light and
antimicrobial combination treatments for surface decontamination of cheese: Favorable and antagonistic
effects. Journal of Dairy Science. 100(3): 1664–1673.
Rajan, S., Pandrangi, S., Balasubramaniam, V. M., and Yousef, A. E. (2006). Inactivation of Bacillus
stearothermophilus spores in egg patties by pressure-assisted thermal processing. LWT – Food Science
and Technology. 39(8): 844–851.
Ramos-Villarroel, A., Aron-Maftei, N., Martín-Belloso, O., and Soliva-Fortuny, R. (2014). Bacterial
inactivation and quality changes of fresh-cut avocados as affected by intense light pulses of specific
spectra. International Journal of Food Science and Technology. 49(1): 128–136.
Raso, J., Calderon, M. L., Gongora, M., Barbosa-Canovas, G. V., and Swanson, B. G. (1998b). Inactivation of
Zygosaccharomyces bailii in fruit juices by heat, high hydrostatic pressure and pulsed electric fields.
Journal of Food Science. 63: 1042–1044.
Raso, J., Pagan, R., Condon, S., and Sala, F. J. (1998a). Influence of temperature and pressure on the lethality
of ultrasound. Applied and Environmental Microbiology. 64: 465–471.
Raso, J., and Barbosa-Cánovas, G. V. (2003). Nonthermal preservation of foods using combined processing
techniques. Critical Reviews in Food Science and Nutrition. 43(3): 265–285.
Raso, J., Condon, S., and Alvarez, I. (2014). Non-thermal processing|pulsed electric field. In: Encyclopedia of
Food Microbiology, pp. 966–973. Batt, C. A., and Tortorello, M. L., Eds., 2nd edn. Elsevier, Oxford, UK.
Raso, J., Palop, A., Pagan, R., and Condon, S. (1998c). Inactivation of Bacillus subtilis spores by combining
ultrasonic waves under pressure and mild heat treatment. Journal of Applied Microbiology. 85: 849–854.
Rathnayaka, R. M. U. S. K. (2013). Antibacterial effect of malic acid against Listeria monocytogenes,
Salmonella enteritidis and Escherichia coli in mango, pineapple and papaya juices. American Journal
of Food Technology. 8(1): 74–82.
Raviyan, P., Zhang, Z., and Feng, H. (2005). Ultrasonication for tomato pectinmethylesterase inactivation:
effect of cavitation intensity and temperature on inactivation. Journal of Food Engineering. 70: 189–196.
Roberts, C. M., and Hoover, D. G. (1996). Sensitivity of Bacillus Coagulans spores to combinations of high
hydrostatic pressure, heat, acidity and nisin. Journal of Applied Bacteriology. 81(4): 363–368.
Rojas, M. L., Trevilin, J. H., Funcia, E. D. S., Gut, J. A. W., and Augusto, P. E. D. (2016). Using ultrasound
technology for the inactivation and thermal sensitization of peroxidase in green coconut water.
Ultrasonics Sonochemistry. 36: 173–181.
Saboyainsta, L. V., and Maubois, J. L. (2000). Current developments of microfiltration technology in the dairy
industry. Lait. 80: 541–553.
Sala, F. J., Burgos, J., Condón, S., López, P., and Raso, J. (1995). Effect of heat and ultrasound on microorganisms
and enzymes. In: New Methods of Food Preservation, pp. 176–204. Gould, G. W. Eds., London, UK:
Blackie Academic and Professional.
Sale, A. J. H., Gould, G. W., and Hamilton, W. A. (1970). Inactivation of bacterial spores by hydrostatic
pressure. Journal of General Microbiology. 60: 323–334.
Salinas-Roca, B., Soliva-Fortuny, R., Welti-Chanes, J., and Martín-Belloso, O. (2016). Combined effect of
pulsed light, edible coating and malic acid dipping to improve fresh-cut mango safety and quality. Food
Control. 66: 190–197.
Santhirasegaram, V., Razali, Z., George, D. S., and Chandran, S. (2015a). Effects of thermal and non-thermal
processing on phenolic compounds, antioxidant activity and sensory attributes of Chokanan Mango
(Mangifera indica L.) juice. Food Bioprocess Technology. 8: 2256–2267.
Santhirasegaram, V., Razali, Z., and Somasundram, C. (2015b). Effects of sonication and ultraviolet-C
treatment as a hurdle concept on quality attributes of Chokanan mango (Mangifera indica L.) juice.
Food Science and Technology International. 21(3): 232–241.
São José, J. F. B., and Vanetti, M. C. D. (2012). Effect of ultrasound and commercial sanitizers in removing
natural contaminants and Salmonella enterica Typhimurium on cherry tomatoes. Food Control. 24: 95–99.
Sara, S., Martina, C., and Giovanna, F. (2014). High pressure carbon dioxide combined with high power
ultrasound processing of dry cured ham spiked with Listeria monocytogenes. Food Research
Sasagawa, A., Yamazaki, A., Kobayashi, A., Hoshino, J., Ohshima, T., Sato, M., Fujii, T., and Yamada, A.
(2006). Inactivation of Bacillus Subtilis spores by a combination of hydrostatic high-pressure and pulsed
electric field treatments. The Review of High Pressure Science and Technology. 16(1): 45–53.
Sengulf, M., Erkaya, T., Başlar, M., and Ertugay, M. F. (2011). Effect of photosonication treatment on
inactivation of total and coliform bacteria in milk. Food Control. 22: 1803–1806.
Setlow, B., Atluri, S., Kitchel, R., Koziol-Dube, K., and Setlow, P. (2006). Role of dipicolinic acid in resistance
and stability of spores of Bacillus subtilis with or without DNA-protective alpha/beta-type small acid-
soluble proteins. Journal of Bacteriology. 188(11): 3740–3747.
Setlow, B., Korza, G., Blatt, K. M. S., Fey, J. P., and Setlow, P. (2016). Mechanism of Bacillus subtilis spore
inactivation by and resistance to supercritical CO2 plus peracetic acid. Journal of Applied Microbiology.
120(1): 57–69.
Schaffner, D. F., Hamdy, M. K., Toledo, R. T., and Tift, M. L. (1989). Salmonella inactivation in liquid whole
egg by thermoradiation. Journal of Food Science. 54: 902–905.
Shahbaz, H. M., Yoo, S., Seo, B., Ghafoor, K., Kim, J. U., Lee, D., and Park, J. (2016). Combination of
TiO2-UV photocatalysis and high hydrostatic pressure to inactivate bacterial pathogens and yeast in
commercial apple juice. Food and Bioprocess Technology. 9: 182–190.
Shamsudin, R., Adzahan, N. M., Yee, Y. P., and Mansor, A. (2014). Effect of repetitive ultraviolet irradiation
on the physico-chemical properties and microbial stability of pineapple juice. Innovative Food Science
and Emerging Technologies. 23: 114–120.
Sharma, G. (2014). Ultraviolet light. In: Encyclopedia of Food Microbiology, pp. 665–671. Batt, C. A., and
Tortorello, M. L., Eds., 2nd edn. Elsevier: Amsterdam, the Netherland.
Shayanfar, S., Chauhan, O. P., Toepfl, S., and Heinz, V. (2014). Effect of non-thermal hurdles in extending
shelf life of cut apples. Journal of Food Science and Technology. 51(12): 4033–4039.
Shimada, K., and Shimahara, K. (1991). Decrease in high pressure tolerance of resting cells of Escherichia coli
K-12 by pretreatment with alternating current. Agricultural and Biological Chemistry. 55: 1247–1251.
Siemer, C., Toepfl, S., and Heinz, V. (2014). Inactivation of Bacillus subtilis spores by pulsed electric fields
(PEF) in combination with thermal energy II. Modeling thermal inactivation of B. subtilis spores during
PEF processing in combination with thermal energy. Food Control. 39(1): 244–250.
Sizer, C. E., and Balasubramaniam, V. M. (1999). New intervention processes for minimally processed juices.
Food Technology. 53: 64–67.
Slieman, T. A., and Nicholson, W. L. (2000). Artificial and solar UV radiation induces strand breaks and
cyclobutane dimers in Bacillus subtilis spore DNA. Applied and Environmental Microbiology. 66: 199–205.
Smith, K., Mittal, G. S., and Griffiths. M. W. (2002). Pasteurization of milk using pulsed electrical field and
antimicrobials. Journal of Food Science. 67(6): 2304–2308.
Spilimbergo, S., Cappelletti, M., Tamburini, S., Ferrentino, G., and Foladori, P. (2014). Partial permeabilisation
and depolarization of Salmonella enterica Typhimurium cells after treatment with pulsed electric fields
and high pressure carbon dioxide. Process Biochemistry. 49: 2055–2062.
Spilimbergo, S., Dehghani, F., Bertucco, A., and Foster, N. R. (2003). Inactivation of bacteria and spores by
pulse electric field and high pressure CO2 at low temperature. Biotechnology and Bioengineering. 82(1):
118–125.
Stewart, C. M., Dunne, C. P., Sikes A., and Hoover, D. G. (2000). Sensitivity of spores of Bacillus subtilis
and Clostridium sporogenes PA 3679 to combinations of high hydrostatic pressure and other processing
parameters. Innovative Food Science and Emerging technologies. 1(1): 49–56.
Su, Y., Zhang, Q. H., and Yin, Y. (1996). Inactivation of Bacillus subtilis spores using high voltage pulsed
electric fields. IFT Annual Meeting Book of Abstracts, Session 26A-14.
Sung, H. J., Song, W. J., Kim, K. P., Ryu, S., and Kang, D. H. (2014). Combination effect of ozone and heat
treatments for the inactivation of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria
monocytogenes in apple juice. International Journal of Food Microbiology. 171: 147–153.
Timson, W. J., and Short, A. J. (1965). Resistance of microorganisms to hydrostatic pressure. Biotechnology
and Bioengineering. 7: 130–159.
Tiwari, B. K., O’Donnell, C. P. O., Patras, A., Brunton, N., and Cullen, P. J. (2009). Stability of anthocyanins
and ascorbic acid in sonicated strawberry juice during storage. European Food Research Technology.
228(5): 717–724.
Tomadoni, B., Cassani, L., Ponce, A., Moreira, M. R., and Agüero, M. V. (2016). Optimization of ultrasound,
vanillin and pomegranate extract treatment for shelf-stable unpasteurized strawberry juice. LWT – Food
Ukuku, D., and Geveke, D. J. (2010). A combined treatment of UV-C-light and radio frequency electric field
for the inactivation of Escherichia coli K-12 in apple juice. International Journal of Food Microbiology.
138(1–2): 50–55.
Unal, R., Kim, J. G., and Yousef, A. E. (2001). Inactivation of Escherichia coli O1 57: H7, Listeria
monocytogenes, and Lactobacillus leichmannii by combinations of ozone and pulsed electric field.
Journal of Food Protection. 64(6): 777–782.
USDA. (2000). US Food and Drug Administration Report. Kinetics of Microbial Inactivation for Alternative
Food Processing Technologies: Ultrasound. Published June 2, 2000. Available at http://www.vm.cfsan.
fda.gov/~comm/ift-us.html.
USDA. (1997). Code of Federal Regulations, Title 9, Part 381.66-poultry products; temperatures and chilling
and freezing procedures. Office of the Federal Register National Archives and Records Administration,
Washington, DC.
Vercet, A., Lopez, P., and Burgos, J. (1997). Inactivation of heat-resistant lipase and protease from Pseudomonas
fluorescens by manothermosonication. Journal of Dairy Science. 80: 29–36.
Vercet, A., Lopez, P., and Burgos, J. (1999). Inactivation of heat-resistant pectinmethylesterase from orange by
manothermosonication. Journal of Agriculture and Food Chemistry. 47: 432–437.
Walkling-Ribeiro, M., Noci, F., Cronin, D. A., Lyng, J. G., and Morgan, D. J. (2009). Shelf life and sensory
evaluation of orange juice after exposure to thermosonication and pulsed electric fields. Food and
Bioproducts Processing. 87: 102–107.
Walkling-Ribeiro, M., Noci, F., Cronin, D. A., Riener, J., Lyng, J. G., and Morgan, D. J. (2008). Reduction
of Staphylococcus aureus and quality changes in apple juice processed by ultraviolet irradiation, pre-
heating and pulsed electric fields. Journal of Food Engineering. 89: 267–273.
Walkling-Ribeiro, M., Rodríguez-González, O., Jayaram, S., and Griffiths, M. W. (2011). Microbial inactivation
and shelf life comparison of “cold” hurdle processing with pulsed electric fields and microfiltration, and
conventional thermal pasteurisation in skim milk. International Journal of Food Microbiology. 144:
379–386.
Walter, R. P., Morris, J. G., and Kell, D. B. (1987). The roles of osmotic stress and water activity in the inhibition
of the growth, glycolisis and glucose phosphotransferase system of Clostridium pasteurianum. Journal
of General Microbiology. 133: 259–266.
Wekhof, A. (2000). Disinfection with flash lamps. PDA Journal of Pharmaceutical Science and Technology.
54: 264–276.
Whangchai, K., Saengnil, K., and Uthaibutra, J. (2006). Effect of ozone in combination with some organic
acids on the control of postharvest decay and pericarp browning of longan fruit. Crop Protection. 25:
821–825.
Williams, R. C., Sumner, S. S., and Golden, D. A. (2004). Survival of Escherichia coli O157: H7 and
Salmonella in apple cider and orange juice as affected by ozone and treatment temperature. Journal of
Food Protection. 67: 2381–2386.
Williams, R. C., Sumner, S. S., and Golden, D. A. (2005). Inactivation of Escherichia coli O157: H7 and
Salmonella in apple cider and orange juice treated with combinations of ozone, dimethyl dicarbonate,
and hydrogen peroxide. Journal of Food Science. 70: M197–M201.
Wong, E., Vaillant-Barka, F., and Chaves-Olarte, E. (2012). Synergistic effect of sonication and high osmotic
pressure enhances membrane damage and viability loss of Salmonella in orange juice. Food Research
Wouters, P. C. and Smelt, J. P. P. (1997). Inactivation of microorganisms with pulsed electric fields: potential
for food preservation. Food Biotechnology. 11: 193–229.
Wu, T., Yu, X., Hu, A., Zhang, L., Jin, Y., and Abid, M. (2015). Ultrasonic disruption of yeast cells: Underlying
mechanism and effects of processing parameters. Innovative Food Science and Emerging Technologies.
28: 59–65.
Xu, W., Chen, H., Huang, Y., and Wu, C. (2013). Decontamination of Escherichia coli O157: H7 on green
onions using pulsed light (PL) and PL-surfactant-sanitizer combinations. International Journal of Food
Xu, W., Chen, H., and Wu, C. (2015). Application of pulsed light (PL)-surfactant combination on inactivation
of Salmonella and apparent quality of green onions. LWT – Food Science and Technology. 61: 596–601.
Yogesh, K. (2016). Pulsed electric field processing of egg products: A review. Journal of Food Science and
Technology. 53(2): 934–945.
Yoo, S., Ghafoor, K., Kim, J. U., Kim, S., Jung, B., Lee, D., and Park, J. (2015). Inactivation of Escherichia
coli O157: H7 on orange fruit surfaces and in juice using photocatalysis and high hydrostatic pressure.
Journal of Food Protection. 78(6): 1098–1105.
Yuk, H. G., Mee, Y. Y., Yoon, J. W., Marshall, D. L., and Deog-Hwan, O. (2007). Effect of combined ozone
and organic acid treatment for control of Escherichia coli O157: H7 and Listeria monocytogenes on
enoki mushroom. Food Control. 18: 548–553.
Yuk, H. G., Yoo, M. Y., Yoon, J. W., Moon, K. D., Marshall, D. L., and Oh, D. H. (2006). Effect of combined
ozone and organic acid treatment for control of Escherichia coli O157: H7 and Listeria monocytogenes
on lettuce. Journal of Food Science. 71: 83–87.
Zhao, W., Yang, R., Tang, Y., and Lu, R. (2007). Combined effects of heat and PEF on microbial inactivation
and quality of liquid egg whites. International Journal of Food Engineering. 3(4): 1–20.
ChaptEr 18
Non-thermal processing of Seafoods
K. Sarika and J. Bindu
CONtENtS
18.1 Need for Non-thermal Preservation Techniques .............................................................. 374

18.2 High-Pressure Processing ................................................................................................. 375
18.3 Mechanism of Pressure Treatment ................................................................................... 375
18.4 Major Advantages of the Technology ............................................................................... 376
18.5 Application of High Pressure in Food Preservation ......................................................... 376
18.6 Fish and Seafood ............................................................................................................... 376
18.7 Shelf Life Extension.......................................................................................................... 378
18.8 Effect of HPP on Microbial Inactivation .......................................................................... 379
18.9 Effect of HPP on Gelation ................................................................................................ 380
18.10 Effect of HPP on Lipid Oxidation .................................................................................... 382
18.11 Effect of HPP on Enzymes Affecting Quality.................................................................. 382
18.12 Effect of HPP on Colour ................................................................................................... 383
18.13 Effect of HPP on Texture .................................................................................................. 383
18.14 Pressure-Assisted Thermal Processing ............................................................................. 384
18.15 Use of Pulsed Light in Seafoods ....................................................................................... 385
18.16 Ultrasound Processing ...................................................................................................... 386
18.17 Conclusion......................................................................................................................... 387
References ...................................................................................................................................... 387
Changes in consumer’s desires in the recent past have led to the requirement for more convenient
foods having supreme qualities and freshness, minimally processed and packaged, easy-to-consume,
and nutritionally healthier. Hence, the focus of food scientists and engineers has been directed towards
alternative technologies or minimal processing and preservation technologies that are environmen-
tally friendly, low in cost, and able to preserve fresh quality attributes of the food. Many novel non-
thermal technologies like high-pressure processing (HPP), pulsed light (PL), ultrasound, irradiation,
etc. find application in preservation of food and are in the line of commercialization. A major trend was
to employ these techniques in newer combinations that effectively do preservation rather exploiting a
single technique to a maximum, commonly called “Hurdle Technology” (Leistner, 1995). In contrast
to the existing technologies, new method of preservation aims at inactivation of microorganisms rather
than inhibiting them. Another field of application is in the development of new and improved products
by functional modification of the food macromolecules that help the processor to have outstanding
quality, with reduced cost, time, and energy. The new technologies now introduce more possibilities in
non-thermal or mild heat alternatives to the conventional heat processing (Gould, 2000).
373
18.1 NEED FOr NON-thErMaL prESErVatION tEChNIQUES
Till last century, food processing and preservation operations worked only on unit operations
involving heat such as thermal pasteurization and thermal sterilization for the inactivation of microor-
ganism and reduction of enzyme activity, that severely affected the food quality. This has resulted in
making safer products with extended shelf life over their raw counterparts. But despite its substantial
benefits, thermal treatments end with over-processed food having significant changes that can alter its
sensorial attributes like flavour, colour, texture, and nutrient content (Barbosa-Canovas and Bermudez-
Aguirre, 2010). This paved way to consumer demand for the fresh like characteristics in their food
having a superior sensory and nutritional quality compared to the existing products in market. Now
consumers are more aware of the contents of food and the technologies used for processing and show
a high preference for natural products (Evans and Cox, 2006). This has become a challenge for the
food scientists and food engineers around the world and led to diversion of focus in new areas of non-
thermal technologies, where different physical hurdles are being explored for microbial inactivation,
while retaining the fresh attributes. During the time HPP has turned to be an explored technology and
today it is a commercial reality. HPP products find place in the world food market with high-quality
and high-value addition. Today, the use of high pressure (300–700 MPa) for commercial application
comes in vessels ranging 35–420 L capacity which had given an annual production of >150,000 tons
(Wan et al., 2009). Regulatory agencies like FDA have approved HPP as substitute to pasteurization,
but in February 2009 a combination of pressure with heat called as pressure-assisted thermal steriliza-
tion (PATS) found to be more effective instead of conventional sterilization (NCFST, 2009).
The new technologies like HPP, pulse electric field, pulse intense light pulses (ILP), X-rays,
and ultrasound have been introducing new scientific approaches towards food processing and
preservation. Among all non-thermal technologies, HPP offers promising possibilities for the
processing and preservation especially in meat, poultry, and seafood.
The first line of HPP was demonstrated in 1899 by Bert H. Hite, as a possible food preservation
process at West Virginia Agricultural Experimental Station (Hoover et al., 1989; Knorr, 1999).
Hite, in his research, observed high-pressure application can delay the souring of milk at ambient
temperatures (Knorr, 2003). Later, Bridgman in 1914 observed coagulation of egg white with high-
pressure treatment. Because of this, after 1914 pressure application has been disregarded mainly
due to absence of general idea on the pressure effect on protein and on biochemical reactions. The
non-availability of suitable high-pressure equipment has ended this technology to receive a wide
recognition (Rastogi et al., 2007), but it re-surfaced in the food industry in the late 1980s. Later in
1982–1988, Hite’s work was repeated by Daniel Farkas, Dallas Hoover, and Dietrich Knorr at the
University of Delaware using a cold isostatic press and revealed the effect of pressure of 350 MPa
(50,000 psi) can inactivate a wide range of pathogenic and spoilage microbes. During the same
period, studies were undertaken in Japan on preservation of foods by high pressure.
In 1992, commercialized high-pressure-processed products (high-acid products including apple,
strawberry, and pineapple jams) were marketed in Japan (Hayashi, 2002). Since then, HPP has been
applied to fruit preserves, raw squid, grape juice, and mandarin orange juice in Japan (Hayakawa
et al., 1996; Rastogi et al., 2007). High-pressure-processed foods are available in the markets of
Japan (Suzuki, 2002) Europe, and United States since 1996 (Knorr, 1999).
In the USA, the impetus for high-pressure technology came from the U.S. Army Research
Center in Natick, Massachusetts, in order to develop better quality MREs (Meals Ready-to-Eat)
for the troops. Collaborations between the University of Delaware and the Oregon State University
lead to successful demonstration of preservation of spaghetti with meat sauce, Spanish rice, yogurt
with peaches and a fruit mix, using HPP. Samples were shown to be microbiologically stable for up
to 120 days at room temperature (Farkas, 2007). The first commercial high-pressure product in the
U.S. was Avo Classic Guacamole (a heat-sensitive product) with extended refrigerated shelf life and
manufactured by Avomex, Inc., Keller, TX. Other commercially available high-pressure-processed
non-tHerMAL ProCessinG oF seAFooDs 375
products in Australia, Europe, and the U.S. include juices, tomato salsa, smoothies, fruit and
vegetable purees, and ready-to-eat meats.
Later, there was a growing interest in the area of seafood safety that led seafood processors to
explore high-pressure technology in product development and extension of shelf life. This technology
was utilized in the area of extending shelf life of product mainly by destroying the spoilage and
pathogenic microorganisms (Toepfl et al., 2006) and also used as an alternative thermal treatment to
packaged food materials. This non-thermal preservation technique could also show many benefits
like complete separation of meat from shell of clams, crabs, lobsters, and oysters providing high
yield of product without any mechanical damage. This technology could open up the new areas
of product development and product improvements in all segments of the meat and fish industry.
Another advent is pressure-assisted freezing and thawing, which finds its unique application in food
industry especially product development and product quality improvement (Lebail et al., 2002).
Since HPP has minimal detrimental impact on thermally labile bioactive compounds, the technology
is becoming a topic of major interest for cosmetic, nutraceutical, and pharmaceutical industry.
18.2 hIGh-prESSUrE prOCESSING
The basic principles that govern the high-pressure effect on the behaviour of foods are
(1) Pascal’s isostatic principle and (2) Le Chatelier’s principle. According to Pascal’s isostatic prin-
ciple high pressure acts uniformly and instantly throughout the sample, independent of the size and
shape of the food product (Smelt, 1998). A uniform pressure will be applied to the product from
all direction, so the product will not get damaged and return to its original shape on the release of
pressure (Balasubramaniam et al., 2008). Isostaticity, i.e., uniform pressure distribution, in a food
subjected to HPP is valid for homogeneous, isotropic foods, but it may not hold for heterogeneous
and unisotropic food products such as meats containing bones and muscle fibres. This issue of pres-
sure non-uniformity remains unresolved and is an active research area.
The fundamental principles of physico-chemical changes occurring during HPP follow the Le
Chatelier’s principle, which states that “when a system at equilibrium is disturbed, the system then
responds in a way that tends to minimizes the disturbance.” So, at high pressure any reaction like
change in conformation, or transition of phase that accompanied volume decrease will be favoured,
while inhibiting those reactions involving an increase in volume (Lopez-Malo et al., 2000). The effect
of pressure and temperature on kinetics is antagonistic. So, temperature effects on both internal energy
and volume of the system in contrast to the effect of pressure on the volume alone. HPP resulted in
small amount of heat due to compression, a thermodynamic outcome and many food and non-food
materials, which are incompressible at atmospheric and low pressures, get compressed under very
high pressure. The work done to compress a material under pressure gets converted into heat which
increases the temperature of the substance. This is called adiabatic heat of compression and is usually
expressed as temperature rise per 100 MPa increase in pressure. Typical values of adiabatic heat of
compression are 3°C for water, 6°C–9°C for cooking oils and fatty foods, and 30°C for synthetic
chemicals such as hexane. For most foods containing water, carbohydrates, fats and protein, the range
of values for adiabatic heat of compression is 3°C–6°C. Therefore, although HPP is considered as a
non-thermal process, in some situations pressure increase will result in significant temperature rise.
18.3 MEChaNISM OF prESSUrE trEatMENt
Each processing cycle consists of an initial pressurization period where the pressure builds up and
the processing operation can be done either with or without the application of heat. The packaged prod-
uct should be in flexible or semi-flexible pouch that can sustain very high pressures. The product is then
submerged into a pressure-transmitting fluid, where water is commonly used. Other liquids like ethanol
or glycol, castor oil, silicone oil, etc. can also be used in various combinations with water or separately.
This fluid is able to protect the inner vessel from being corroded and the fluid is selected based on the
manufacture’s specification. During the pressure processing adiabatic heating occur and the product
gets heated. The temperature increase due to adiabatic heating depends on the type of fluid, pressuriza-
tion rate, temperature, and pressure. In the case of water, the increase in temperature due to adiabatic
heating is 3°C for every 100 MPa increase in pressure, which indirectly affects the process temperature.
Once the process starts, the hydraulic fluid is pressurized with a pump and the generated pressure
is transmitted into the packaged food uniformly from all sides. Since this processing is independent
of size and geometry of foods, also acts instantaneously the total processing time can be reduced. The
process is suitably applied to liquid foods and other foods having a certain amount of moisture content.
The transmitted pressure is uniform and simultaneously applied from all directions, therefore the food
retains its structure even at high pressures. Once the pressure is built up to the desired level the product
is held at this pressure for a few minutes and then decompression or pressure release takes place. Once
there is a fall in pressure the product temperature falls below that of the initial product temperature.
18.4 MaJOr aDVaNtaGES OF thE tEChNOLOGY
1. HPP does not involve in breaking covalent bonds which prevents the development of unpleasant
flavours to the product and maintains the natural freshness and quality.
2. High pressure is able to modify the palatability and functional properties by inducing denaturation
and muscle protein gelation.
3. Process can be carried out at ambient temperatures, which helps in reducing the thermal energy
used during conventional processing.
4. HPP is isostatic in nature, equally applied to all particles of food, with no particle escapes.
5. Since high pressure is not time-mass dependent, pressure acts instantaneously thereby reducing the
processing time.
6. This non-thermal technology is independent of size and geometry of the food.
7. The process is eco-friendly, with no waste and requires only electric energy.
18.5 appLICatION OF hIGh prESSUrE IN FOOD prESErVatION
Food, a proficient medium for the growth and multiplication of bacteria. So, the processing opera-
tions should be able to inactivate this microorganism especially spoilage and pathogenic bacteria.
Pasteurization is employed for high-acid foods and sterilization for low-acid foods for the inactivation
of both microorganisms along with the reduction in enzymatic spoilage. Thermal treatments can-
not maintain the original nutrient content, the structure and functionality of the ingredients. High-
pressure-processing technology has shown a negligible effect on the nutrient content of food and has
interesting functional effects. High pressure can able to preserve the fresh sensory attributes of the
food unlike in thermal processing, where heat works as a catalyser for some chemical reactions involv-
ing oxidation, browning, protein denaturation etc. Since pressurization had any effect on the covalent
bonds, reactions involving developing strange off-flavour in food are avoided (Knorr, 2002).
18.6 FISh aND SEaFOOD
The advent of HPP technique in food industry has been extensively utilized in shelf life exten-
sion by ensuring microbiological safety and quality in different foods. The effect of pressure on
fish and seafood has not been as intensively studied as this technology had a wide adoption in other
industries dealing with food products like juice, cooked meat, and dry-fermented sausages. Since
the fish and shell fishes are highly perishable, the processing and preservation techniques to inhibit/
retard spoilage and extend the shelf life would be an added advantage (Rode and Hovda, 2016). The
highly perishable nature of seafoods spoil them faster than other muscle foods and hence they are
more susceptible to post-mortem texture changes than other muscle foods.
HPP-related studies of fish comprised the researches from Campus (2010), Ramirez-Suarez
and Morrissey (2006), and Yagiz et al. (2009). Another practical problem is in the difficulty of
comparing many studies due to the various equipment models with different ranges of pressure
levels, holding time, temperature, storage time analysis, etc. and different biological composition
of fishes and shelf life (Rode and Hovda, 2016). The fish quality depends on the various changes
involving rapid breakdown processes which resulted in the quality losses. Oxidation of lipid and
degradation of microbial quality are the major two quality loss factors. Seafood’s especially bivalve
shellfishes are more featured for food-borne diseases (Potasman et al., 2002), probably because of
the filter feeding mechanism that selectively accumulates bacteria and viruses from the surrounding
waters (Richards, 1988). The shell fishes are likely to be eaten as raw including the intestinal tract,
or sometimes mild heat-treated (Gram and Huss, 2000). The inactivation can be achieved through
the thermal treatments, which loses the natural taste and appearance of shellfish, leading to non-
acceptance by the consumers. This loss of natural sensory attributes can be overcome by the use
of HPP where the nutritional qualities are retained as in unprocessed product (Hoover et al., 1989;
Smelt, 1998). This novel technique had a wide range of application especially in improving the
microbial, physico-chemical, and sensorial qualities. The application of high pressure shows a great
potential in improving the physico-chemical, microbial, and sensory quality of fish muscles. The
inactivation of microorganism and the ability to retard/inhibit autolytic enzyme activities led to the
shelf life extension in fish products. The fish quality changes are mainly contributed by bacterial
growth and multiplication but the changes occurred in tissues are mainly resulted from autolytic
reactions controlled by native enzymes like ATP break-down process (Kennish and Kramer, 1986)
which in turn affect the quality. Fish is characterized by the presence of odourless compounds called
trimethylamine oxide (TMAO), which on spoilage is converted to trimethylamine by bacterial
enzymes and is used in the assessment of quality. Generally volatile bases are produced in fish
muscle by autolytic enzymes, putrefactive micro-organisms or by chemical reactions.
High pressure has been found to inhibit the formation of putrefactive compounds and maintains
the hardness of fish muscles during storage, thereby, ensuring higher sensory quality compared to
untreated ones. However, at higher pressures the formation of cooked appearance, discoloration
and lipid oxidation are the main setbacks that could limit the application of this technology on fish
muscles. Pressure can be used to eliminate pathogens like E. coli, Salmonella, Listeria, and spoilage
bacteria without significantly affecting colour and flavour of the product. Thus, this technique can
play a vital role in ensuring microbial safety, thereby extending the shelf life of fish without heat.
Another line of research is in the development of TTI-Enzymatic time-temperature integrators,
as an intrinsic biological marker for assessing the heat-induced quality changes or inactivation of
bacteria (Rode and Hovda, 2016). Research has been going on in developing TTIs for HPP (Grauwet
et al., 2012). The studies showed that a pressure above 300 MPa can cause a reduction of initial
bacterial load/growth of spoilage microorganism and enzymatic activity in many fish products
stored under chilled conditions (Yagiz et al., 2009; Erkan et al., 2010; Kamalakanth et al., 2011).
The utilization of HPP in the field of shell fish processing attracted worldwide attention,
especially for the complete removal of meat from shell and also to provide a safe mechanism for
raw seafood consumption. The application of HPP in muscle foods is either for tenderization of
the muscle or for extension of shelf life. HP can be used to modify functional properties of the
food material while simultaneously enhancing safety of raw seafoods and retaining its sensory and
nutritional qualities (Cheftel and Culioli, 1997). An interesting field is the development of new gel-
based products with consumer-desired sensory attributes and mouthfeel and many researches are
diverted in this area. Another promising area for development of shelf-stable ready-to-eat products
is pressure-assisted thermal processing as an alternative to the high-temperature conventional
processing. Besides this, pressure-induced gelling and pressure-assisted freezing/thawing, which
helps in retaining the microstructure and reduced drip losses, are the focusing areas in fish
processing.
18.7 ShELF LIFE EXtENSION
1. Fish
Fish being a low-acid food requires chilled temperatures for extension of shelf life as pressure
process cannot ensure a complete sterility. So pressurized fish products should be always stored at
chilled temperatures. Several studies on shelf life extension of different fish have been conducted. An
extended shelf life of more than 22 and 93 days was observed in HP-treated albacore tuna stored at
4°C and −20°C, respectively (Ramirez-Suarez and Morrissey, 2006). HP-treated abalone was extended
more than 65 days of shelf life irrespective of HP treatment applied as control (30 days). Different
HP treatments were applied to establish the best processing conditions for the quality and extension
of shelf life of red mullet stored at 4°C (Nuray et al., 2010) and a shelf life of 12 days for untreated
red mullet and 14 and 15 days for treated red mullet at 220 MPa for 5 min at 25°C and 330 MPa for
5 min at 3°C was obtained, respectively. High-pressure-treated sliced raw squids had a high reduction
in the psychrophilic count. Pressure level of 220 MPa and a 30 min holding time were optimal and
most effective in prolonging the storage period of tuna muscle (up to 9 days), as well as in reducing
the proteolysis activity, texture degradation, TVBN, and histamine formation (Zare, 2004). Ramirez-
Suarez and Morrissey (2006) reported an enhancement of shelf life of minced albacore muscle for
more than 22 days at 4°C and more than 93 days at −20°C when high-pressure-processed at 275 and
310 MPa. They observed a low microorganism level, colour change of the muscle, and induced the
formation of high-molecular-weight polypeptides most likely through disulphide bonding, thereby
promoting texture improvement in minced albacore muscle. HPP in combination with salting and
smoking showed an enhanced effect on shelf life than individual effect. (Montero et al., 2007).
Similar effect was noticed in the combination effect of high pressure and short treatment in improving
the quality of smoked salmon (Gudbjornsdottir et al., 2010). High pressures of 700 MPa has been
successfully used to extend the shelf life of salmon spread product to 180 days (Carpi et al., 1995).
Another area of application is in marination and impregnation of desired flavours and colours with
high pressure will also pave its way to the introduction of new consumer attractive products in market.
2. Shellfish
Oysters are commercially produced in different countries, especially in the USA. Besides microbial
preservation and shelf-life extension, high-pressure treatment helps in shucking of oyster meat out
of their shells, which is helpful. A 5-log reduction of target microorganisms in oysters was noticed
when high pressurized at 400 MPa and stored for 41 days at 2°C (Lopez-Caballero et al., 2000b).
A comparison on physical and biochemical changes in oysters was done by Cruz-Romero et al.
(2007) when subjected to high-pressure treatment (260 MPa for 3 min) and heat treatment (cool
pasteurization (CP) at 50°C for 10 min and traditional pasteurization (TP) at 75°C for 8 min) on the
shucking yield, and reported that HP-treated samples were best among the three, with CP and TP
showing negative effect on the shucking yield.
Patterson et al. (2003) reported that when oyster samples were pressure-treated, a low initial
microbial count was observed and samples did not attain spoilage levels during four weeks of
storage. A sensory acceptance for appearance was significantly more in pressure-treated samples
(>400 MPa) than control and the raw odour did not significantly change up to two weeks of storage.
Many researches were done in the area of shucking of live clam, oysters, and mussels. Live oysters
subjected to moderate pressures of 240–350 MPa for 3 min helped to open up the oysters without
knife and, thus, have developed as an alternative to the laborious and costly hand-shucking process
in oyster industry. Optimum shucking pressures range of 240 to 275 MPa can cause minimum
changes to pacific oyster (He et al., 2002).
Mussel meat is usually consumed raw, blanched, or cooked and meat removal from the shell is
done by steaming or dipping in boiling water. These process leads to the loss of structure, moisture
reduction, protein degradation, and loss of juiciness resulted in rubbery texture. HPP helps in
shucking the raw meat without cooking, so as to remove rigid shell of crustaceans and molluscs
easily without losing the natural texture and appearance (Errol, 2007). Thus, a more efficient means
of meat removal can be achieved without changing the size and shape of the meat and also retain the
nutritional qualities (Bindu et al., 2015). In the U.S. market, Motivatit Seafoods Inc. has launched
HPP-treated oysters as Gold Band Oysters marketed by Nisbet Oyster Company. High-pressure
inactivation of microorganism renders the safe sea food for eating in raw conditions (Lopez-Caballero
et al., 2000b). Application of pressure in the range of 230–586 MPa resulted in a reduction in 6 logs
of Vibrio spp. In raw oysters (Koo et al., 2006). Linton et al. (2003) focused on the detailed study
on the effect of high pressure on bacterial flora of oysters, mussels, prawn, and scallops. Narwankar
et al. (2011) did a study on the clams treated under high pressure and observed a reduction of total
plate count (TPC). Patterson et al. (2003) had studied the effect of pressure-treated mussel samples
at 500 MPa or higher and stored under 2°C. They observed an extension of shelf-life of 14 days,
when spoilage level of psychotropic count was assumed to be 106 –107 cfu/mL. He also stated that
fresh and untreated samples were most acceptable, while 600 MPa-treated mussels were acceptable
after 2 weeks of storage based on the sensory analysis of the cooked mussels. Bindu et al. (2015)
found a significant effect on the shucking of meat from the shell by high pressure, together with
shelf life extension of green mussel meat during chill storage. Higher pressures of 300 and 400 MPa
can cause denaturation of the adductor muscles and helped in detachment of meat from the shell.
Pressure-processed green mussel have an extended shelf life of 28 days under chilled storage. Even
though mussels are exploited commercially, the work done on the preservation of meat quality of the
mussels by application of high pressure is relatively less.
Kaur et al. (2013) reported an extension of 15 days’ shelf life in high-pressure-treated tiger
shrimp at 435 MPa as compared to 5 days in untreated sample. The vacuum packed and pressure
treatment of 200 and 400 MPa lead to further shelf life extension of 21 days and 35 days, respectively,
compared to 14 days for untreated shrimp samples (Lopez-Caballero et al., 2000a). Ginson et al.
(2013) observed a trivial reduction in total volatile compounds, total enterobacteriaceae count, and
K-value after HP treatment for 5 min in headless shell on Indian white prawn and found a better
sensory acceptability in samples treated at 270 MPa.
18.8 EFFECt OF hpp ON MICrOBIaL INaCtIVatION
The high-pressure inactivation of microorganism without heat has received a great deal of attrac-
tion, especially in the food industry. Many research project have been carried out in this context and
the application of this non-thermal technology has extended in the various levels of food processing
and preservation. The application of very high pressures especially above 200 MPa and above nor-
mal temperatures (45°C) can inactivate most of the vegetative and pathogenic microorganisms and
the inactivation rate depends on the peak pressure (Patterson, 2005; Lau and Turek, 2007). But, the
rate of inactivation was more preferred at higher pressures of 600 MPa in the food industry except
for protein foods, where denaturation should be avoided. Always the pressure resistance of microbes
often observed at normal temperatures. This helps in improving the inactivation process by varying
the process temperature (Sudheer et al., 2011). The mechanism of inactivation of bacteria has been
detailed by many researches. Torres and Velazquez (2005) studied the mechanism and stated that
pressure can cause cell membrane damage and had an effect on the cellular fluid transport mecha-
nism as well as microbial enzyme denaturation. The inactivation depends on various factors such
as type of microbes, food composition, water activity, pH, etc. The pressure resistance was shown
more in gram-positive bacteria than gram-negative ones due to the hard cell wall. The pressure can
cause damage to cell membrane easily and denature the proteins leading to cell death. The water
activity had a significant effect on the pressure resistance of microbial inactivation. The resistance
was noticeably higher when the water activity decreases. So, a pressure temperature combination
may be beneficially for the delivering sub-lethal stress to bacteria and reviving of bacteria after pres-
sure treatment needs to be checked after some time. The difficulty of microbial inactivation dealt
with the spore, which requires high pressure, temperature and holding time. A pressure at and above
1000 MPa is required for the complete destruction of the spores (Cheftel, 1995). Among the patho-
genic spore formers, the Clostridium botulinum strains has been identified as more pressure resis-
tant while Bacillus amylolique faciens notices as non-pathogenic spore former (Margosch et al.,
2006; Ahn et al., 2007). A more detailed research is required in the area of pressure-temperature
kinetics of microorganism and a more comprehensive database of standardized process conditions,
equipment and microbial techniques should be developed (Balasubramaniam et al., 2004).
The inactivation of yeast and mould can be achieved at 300–400 MPa (25°C) in a few minutes,
but the inactivation yeast ascopores requires higher treatment. Viruses are more resistant to pressure
treatment due to their high structural diversity (Smelt, 1998), except some human viruses. The
inactivation pattern of the virus needs extensive studies and this is an area of concern. Another
characteristic of this technique is the generation of heat of compression or adiabatic heating during
the pressurization process, which depends on many factors like amount of pressure, pressure
transmitting fluid, geometry of equipment, etc. The heat generation is not under control in most
cases and the problem associated with this is a non-constant lethality rate, which resulted in
microbiological tailing effect that can confound the analysis of inactivation kinetics (Ting, 2007).
18.9 EFFECt OF hpp ON GELatION
Gelation is the network formation that holds water in less mobile state due to partial denatur-
ation, followed by aggregation of myosin heads through disulphide bonds and helix coil transition
of tails resulting in a 3D structure. Pressure-induced gelation involves head to head interaction
of myosin. The native myosin exists as monomer, with two heads differentiated. When pressure
is applied, myosin undergoes head-head interaction and exists as oligomer. This oligomer gets
compacted and increase in size on storage. The pressure-induced gelation depends on protein and
treatment conditions like how much pressure, holding time and temperature of the process. So,
an adequate pressure and temperature can enhance the elasticity and tenderness of myofibrillar
protein gels (Jimenez Comenero, 2002). Messens et al. (1997) reported that at lower pressures
(210 MPa up to 30 min), myosin still exists as monomers and pressure cannot induce a proper
gelation. This happens because unlike in heat-induced gelation, the lower pressure was not able
to form tail-to-tail interactions in myosin structure, resulting in the formation of weaker gels. But
on further subjection to heat treatment, the same pressure-treated gels showed similar gel rigid-
ity and microstructure as that of heat-induced ones. This states pressure treatment at lower levels
can retain the original helical structures of myosin, unlike in heat treatment, which is thought
to be related to helix-coil transitions. Protein structure is maintained by interactions within the
protein chain and surrounding solvent. High pressure can cause disturbance in these interactions
and lead to denaturation, aggregation, or gelation. The change in these interactions is primarily
related to rupture of non-covalent interactions within protein and subsequent reformation within
and between protein.
The native conformation of proteins is maintained by non-covalent interactions. The pressure
above 150 MPa has been able to disturb these interactions and cause protein denaturation (Masson,
1992). At the same time the hydrogen bonds are pressure insensitive, so that collagen has been
reported to be scarcely affected by high pressure (Gekko and Koga, 1983; Heremans, 1995). High-
pressure treatment around 300–400 MPa could not significantly affect the intramuscular collagen
solubility of beef (Suzuki et al., 1993) or pacific blue whiting (Fernandez-Martin et al., 1998).
However, bovine collagen solubility has been shown to vary with application of pressure; 200 MPa
increased the solubility, whereas 400 MPa reduced it (Kwiatkowska et al., 2001).
The influence of high pressure towards the susceptibility of protein towards other treatments
was supported by Ashie and Lanier (1999). They reported that pressure treatment could enhance
the TGase activity and render myosin more accessible to transglutaminase catalysed cross linking.
Also, neither endogenous and nor added TGase was affected by high pressure up to 300 MPa at
4°C and above which activity of TGase will be lost. The effect of pressure on TGase was more
obvious in case of raw gels. Now the Hydrostatic pressure can be used for inducing gelation of
different kinds of surimi (Hayashi, 1989) from pollack, sardine, skipjack tuna, and squid. Pressure
can also enhance protein gelation in a poorly-solubilized actomyosin complex (Cando et al., 2014).
The gel properties were varied depending on the type of species. The pressure-induced gels were
smoother and more elastic, especially in marine species, than heat-induced ones and were having
organoleptically superior characteristics. Several studies have been reported on UHP technology
to improve the gel-forming ability of sardine muscle. A similar enhancement in thermal gelation
ability was seen in squid meat paste, as high pressure was applied and found that the breaking
strength value of the gel formed by the pressure of 400 MPa was twice as high as that of thermal-
induced gel (Nagashima et al., 1993).
Sareevoravikul et al. (1996a) studied the effect of high pressure on blue fish meat and found
that application of 3000–3740 atm pressure for 30 min can formulate gels from bluefish meat
paste. The pressure-induced gels were compared with heat-induced gels formulated at 90°C for
20 min or 60°C for 60 min for the properties and observed that gels formed by pressure were more
translucent than by heat. The studies on protein digestion and salt extractability indicated that
pressure-induced gels had less protein denaturation and were more digestible and the proteolytic
activity studies showed that the pressure range used in this study had less effect on the integrity
of endogenous proteases than heat. Nagashima et al. (1993) observed that pressure treatment up
to 1000 MPa for 20 min was effective in inducing thermal gelation of squid meat when applied
before heating, thereby improving its lower thermal gelation ability. Another area of application
is in the field of pressure extraction of gelatin from fish skins and it has been found to be a useful
alternative to the conventional procedure. Gomez-Guillen (2005) reported a drastic reduction in
the treatment time in the longest phase of the treatment, thus, making a gelatin of high gelling
quality in few minutes.
The potential application of HPP has been reported for surumi and surimi-based products
(Uresti et al., 2004, 2005, 2006), cold-smoked fish (Lakshmanan et al., 2007), thermal
processing (Ramirez et al., 2009), and pressure-assisted freezing (Alizadeh et al., 2007) and
thawing (Rouille et al., 2002). Kunnath et al. (2015) observed the formation of softer and glossier
gels when subjected to higher pressures at 400, 500, and 600 MPa than heat-induced hard gels
in pink perch sausage. The texture of pressure-induced sausages was softer, cohesive, and
less chewy and gummier than heat-treated ones. Also, these pressure-processed gels exhibited
greater elasticity and more stable networks in protein which happened due to the increase
of physical interactions like hydrogen and hydrophobic bonds and dispersive interactions,
prompting protein gelation (Sun and Holley, 2010; Cando et al., 2014). These interactions can
easily break and recombine, increasing the elasticity and conformational flexibility of the
network (Herranz et al., 2013).
Many reports have shown that pressure application prior to thermal treatments enhanced the
gelation properties of the cooked gels. When pressure of 300 MPa for 60 min at 0°C was applied,
gelation temperature of milkfish paste was found to reduce from 50°C to 30°C and excellent gels
were obtained when cooked at 90°C for 10 min (Ko, 1996). Similarly, Montero et al. (1997) noticed
high pressure of 300 MPa and subsequent heating at 90°C can cause reduction of gelling time in
washed sardine mince and enhanced the rheological properties when compared with heating. When
ground blue whiting muscle was subjected to HPP at non-denaturing temperatures, the proteins
unfolded at a greater rate, with increasing pressures, but at denaturing temperatures, pressure
prevented fish protein from subsequent thermal denaturation (Fernandez-Martin, 1998).
18.10 EFFECt OF hpp ON LIpID OXIDatION
The most important concern during HPP is the hastening of lipid oxidation. Fish is rich in
polyunsaturated fatty acids (PUFA) content that makes it more prone to lipid oxidation. High-
pressure treatment has resulted in higher TBA values (mg malonaldehyde/kg) for pressure-
processed samples when compared to the control samples – the case of cold-smoked salmon
when subjected to 300 MPa pressure (Lakshmanan et al., 2005). This is mainly due to the fact
that during high-pressure treatment there is a possibility for the cell structure to break and make
available intercellular lipid deposits for oxidation. The accelerated lipid oxidation during high
pressure may be due to the auto-oxidation of fat which is promoted by the release of metal ions
from the denaturation of heme-protein (Tanaka et al., 1991). Lipid oxidation was also reported
by Yagiz et al. (2007) in dark muscle of trout at pressure levels beyond 300 MPa, and pressure of
200 MPa for 30 min duration increased oxidation of lipid in turbot and cod muscle (Angsupanich
and Ledward, 1998). The presence of metal ions like Fe and Cu in the fish muscles also enhanced
oxidation. Hultin (1994) suggested that the changes occurring in the cell membrane during high-
pressure treatment may also make fish muscle less susceptible to oxidation even though the heme
proteins are released. Pressure treatment can also bring about changes in the fish so that the anti-
oxidant properties of astaxanthin present in the muscle is more available to retard further oxida-
tion (Shimidzu et al., 1996). A decrease in the lipid oxidation has been observed in dark muscle of
salmon treated at 300 MPa when compared to 150 MPa and cooked salmon (Yagiz et al., 2009).
High pressures in combination with other pre-treatments will create a hurdle for the microbes
and retard the spoilage, thereby extending the shelf life of the product. Similar findings were
marked by Truong et al. (2016) in fish subjected to high pressure by the formation of unwanted
changes such as acceleration of lipid oxidation, discoloration, and cooked appearance. Vazquez
et al. (2013) observed a significant increase of TBA values after HPP at 150–300 MPa for 5 min,
but, later the trend was decreased for pressure above 450 MPa for 5 min. High pressure in the
range of 275 to 310 MPa (10°C) for 2–6 min also showed lower TBA values (Ramirez-Suarez and
Morrissey, 2006).
The positive effect of pressure on the inactivation of oxidative endogenous enzyme before
storage and processing of fish products was described by Murchie et al. (2005). Vazquez et al.
(2013) noticed a reduction of lipid hydrolysis in Atlantic mackerel (S. scombrus) and Atlantic
horse mackerel (Trachurustrachurus) samples when given a pressure treatment before freezing and
frozen storage (Torres et al., 2011). High pressure can also accelerate the FFA formation by altering
the interactions (electrostatic, VanderWaals, hydrogen bonding, and hydrophobic forces) between
myofibrillar proteins and FFA which causes the FFA to release under high pressure (Sequeira-
Munoz et al., 2006). The effect of pressure on lipid oxidation depends on the type of fish muscle
(white/dark), amount of heme proteins, type of fish species and treatment conditions.
18.11 EFFECt OF hpp ON ENZYMES aFFECtING QUaLItY
Enzymes are a special class of proteins in which biological activity arises from an active site,
brought together by the three-dimensional configuration of the molecule. Any small changes in the
active site can lead to the loss of enzyme activity. High pressure causes protein denaturation, which
is associated with conformational changes leading to change in the functionality of the enzyme
(e.g., increase or loss of biological activity, change in substrates specificity).
High pressure can either deactivate or increase the enzymatic activity in food systems which
has been shown in many real and model food systems. At pressure above 200 MPa and elevated
temperatures, a clear reduction in autolytic activity had been observed and the effect was reduced
by step-pulsed than by continuous pressurization (Hurtado et al., 2001).
HPP is increasingly being accepted as a preservation technique of food that can inactivate
spoilage and pathogenic microorganisms and enzymes ensuring a safe and stable product. An
added advantage is that food quality characteristics like flavour and vitamins are unaffected
or only minimally affected during pressure processing at ambient conditions. In contrast, food
quality enzymes get deactivated during the pressure applications. The inactivation of enzymes
strongly depends on the type of enzyme and the amount of pressure applied. Many enzymes can be
deactivated at lower pressures at room temperature, while some can withstand higher pressures of
1000 MPa range. In such cases, a combined process of pressure and temperature might be necessary
for enzyme inactivation.
18.12 EFFECt OF hpp ON COLOUr
Pressure had a significant effect on the colour of fish products. Pressure-treated samples
appeared like cooked fish, due to the loss of transparency. When the pressure and holding dura-
tion increases, the whiteness of the fish increased. HP causes denaturation of the myofibrillar
and sarcoplasmic proteins which leads to the loss of translucency of flesh affecting the fish
colour. The whitening effect observed in the flesh after high-pressure treatment may also be due
to the denaturation of the globin protein and release of the heme (Carlas et al., 1995) as observed
in cod and mackerel subjected to high pressure (Ohshima et al., 1993). When high pressure is
applied, an increase in L* and b* value had been noticed with mostly an unchanged a* value and
a lower hue angle. The change in hue angle in barramundi (Truong et al., 2016) after pressuriza-
tion was similar to pressurized sea bass (Cheret et al., 2005), but showed a higher hue value in
tuna after pressurization (Ramirez-Suarez and Morrissey, 2006). The initial orange pink colour
of the flesh of salmon, which is the most desirable indicator of quality, also decreased after pres-
surization, which may be due to lipid oxidation caused by degradation of the highly unsaturated
carotenoids such as astaxanthin, a major pigment in salmon. Many researchers have reported
the colour change in fish from raw to cooked appearance, which depends on pressure level and
fish species (Master et al., 2000). Ashie and Simpson (1996) described that a pressure beyond
200 MPa could cause lightness in bluefish and sheep headfish. Atlantic salmon is probably one
of the most sensitive fish to high-pressure-induced colour change (Amanatidou et al., 2000;
Yagiz et al., 2009). Similarly, a significant increase in whiteness was noted in carp treated at
140 MPa and 4°C for 15 min (Sequeira-Munoz et al., 2006) and 150 MP and 20°C for 15 min in
tuna samples, but there were no significant differences in red mullet treated up to 220 MPa at
10°C for 15 min (Erkan et al., 2010).
Truong et al. (2016) found that high pressures at or above 200 MPa resulted in increased whiteness
and loss of translucency in barramundi muscles. The fish muscle retained its raw appearance below
150 or 200 MPa, and with 250 MPa or higher, muscle become opaque like milky white, as if it is
cooked. The high-pressure effect on the fish colour was reported in hake (Hurtado et al., 2000), red
mullet (Erkan et al., 2010), mahi-mahi (Yagiz et al., 2007), and tuna (Zare, 2004).
18.13 EFFECt OF hpp ON tEXtUrE
Denaturation of sarcoplasmic and myofibrillar proteins occur during high-pressure treatment.

The hardness may be due to the unfolding of the sarcoplasmic proteins and formation of new
hydrogen-bonded linkages (Angsupanich and Ledward, 1998). The myosin from fish source may
denature by pressure and form a gel-like texture (Cheftel and Culioli, 1997). HPP also affects
contractile proteins especially actin which unfolds at higher pressure (Yoshioka and Yamamoto,
1998). Proteolytic enzymes in fish get activated at low pressures and inactivation happens at higher
pressures, which leads to fish textural changes and make the meat more tender (Ohshima et al.,
1993). Postmortem changes of fish texture are caused due to modifications in myofibrillar proteins,
by both protease action and variation of physical and chemical conditions. TMAO-ase converts
TMAO into dimethyl amine and formaldehyde. When formaldehyde reacts with fish protein it
forms cross-links that result in the reduction of solubility and a decrease in tenderness. This occurs
in the fish muscle during frozen storage when concentration of oxygen is low. Master et al. (2000)
found out a textural change in cod-like fish species during high-pressure treatment. The authors
observed an increase in hardness at 200 and 400 MPa because of pressure influence on the enzyme
activity. High pressure can cause modification of hydrogen and hydrophobic bonds that brings
about changes in protein structure which in turn alters water bond, protease activity, myosin gel
formation, and sarcoplasmic proteins (Heremans, 1982; Anguspanich and Ledward, 1998).
Truong et al. (2016) found a significant increase in the hardness of barramundi muscle when
pressurized at 150–200 MPa, at the same time hardness was decreased when pressure-treated
at 250 and 300 MPa. A similar trend of observation was earlier reported by Ashie et al. (1997)
on bluefish muscle high pressurized at 300 MPa for 30 min at room temperature. The reason is
that unlike in mammalian muscle, post mortem tenderization happened, which is one of the most
unfavourable quality changes in fish muscle that results in muscle softening and gaping (Cheret
et al., 2007; Terova et al., 2011). The pressure treatment always resulted in the hardness of the fish
muscle (Zare, 2004; Ashie et al., 2007; Yagiz et al., 2009). The springiness increased during storage
in pressurized fish muscle which could be due to the formation of pressure-induced hydrogen-
bonded networks and protein-protein interactions (Truong et al., 2016).
The drip is an important aspect, especially in frozen stored samples, that had a significant
influence on the sensory quality of the fish foods. With increase in pressure, the drip also
increased significantly which can be seen in carp (Wada and Ogawa, 1996), hake (Hurtado
et al., 2000), salmon (Lakshmanan et al., 2007), and tuna (Zare, 2004). Reason might be due to
the structural modification of myofibril protein, the principal site of water retention in muscle
(Offer and Trinick, 1983; Xiong, 1997) and conformational change in the protein mainly due
to disruption of electrostatic and hydrophobic interactions (Zare, 2004; Ramirez-Suarez and
Morrissey, 2006).
18.14 prESSUrE-aSSIStED thErMaL prOCESSING
The pressurization at elevated temperatures has been appraised as an alternative to classical

retorting technologies in food industry (Heinz and Knorr, 2002; Koutchma et al., 2004). This high-
pressure thermal sterilization has led to the introduction of a new dimension of thermal preserva-
tion with enhanced product quality. Pressurization process contributes compression heating and
decompression cooling to the product which adds to the lower thermal load applied to the product
in comparison to conventional thermal processes. Many scientific publications have discussed the
application and impact studies of the high pressure and high temperature effect on the microor-
ganisms, spores and functional modifications over the last decades (Sale et al., 1970; Gould, 1977;
Cheftel and Culioli, 1997; Heinz and Knorr, 1998, 2002; Setlow, 2003; Margosch et al., 2004;
Matser et al., 2004; Eisenbrand, 2005; Knoerzer et al., 2007; Considine et al., 2008; Mathys, 2008;
Wimalaratne and Farid, 2008; Heinz and Buckow, 2010; Knorr et al., 2011; Mújica-Paz et al., 2011;
Reineke, 2012; Reineke et al., 2013).
The technology of HPP is now widely accepted across the world and the trend in adoption of this
technology is increasing every year especially in juice (beverage) sector. Now the consumers have
shown high acceptance of pressurized foods (Olsen et al., 2010). The high-pressure effect on various
systems still needs to be explored. Not only pressure inactivated the microorganism and spores, it
decreased toxicological potential of a food (Sevenich et al., 2016).
18.15 USE OF pULSED LIGht IN SEaFOODS
The introduction of non-thermal preservation techniques in food processing and preservation

have attracted the attention of world especially the food manufacturers (Palmieri and Cacace,
2005). PL technology is one such explored technology in the food industry. This technique works
by applying high-voltage, high-current short electrical pulse to the inert gas in the lamp, which
results in strong collision between electrons and gas molecules causing excitation of the latter,
which then emit an intense, very short light pulse to decontaminate and sterilize foods (Palmieri
and Cacace, 2005). Usually short pulses of light, one to twenty flashes per second, are used in
food industry. Exposure to PL is in the form of high-intensity UV light pulses resulted in micro-
bial inactivation through the process of photochemical, photo thermal, and photo physical route
(Krishnamurthy et al., 2010). The microbial inactivation is mostly due to the photochemical action
of the ultra-violet part of the light spectrum, which can cause thymine dimerization in the DNA
chain preventing replication and ultimately leading to cell death (Gomez-Lopez et al., 2007). The
lethal effect is caused by the combined action of light pulses on proteins and nucleic acids in the
cell (photochemical effect), a transient temperature increase caused by heat dissipation of light
pulses penetrating the product (photothermal effect) and morphological damage to cells caused by
the constant disturbance originating from the high-energy pulses (photo-physical effect). Hence,
the inactivation of microorganism is achieved in the food, without causing any adverse effect to the
overall characteristics (Palmieri and Cacace, 2005).
Bank et al. (1990) seem to be the first group who published in the scientific literature on the
PL application and its inactivation of microorganism. Recent studies showed the efficacy of PL on
inactivation of microorganisms in several seafoods (Dunn et al., 1995), salmon filets (Ozer and
Demirci, 2006), ready-to-eat sausages (Uesugi and Moraru, 2009), etc. Ozer and Demirci (2006)
reported 1-log reduction of Escherichia coli O157:H7 or Listeria monocytogenes in PL-treated
salmon fillets at fluence of 5.6 J cm2. The application of PL has been conducted in various foods
but only few researches have been reported in fish and fishery products. Takeshita et al. (2003)
found that microbial inactivation can be dealt with the damage of membranes, proteins, and other
macromolecules within microorganisms when PL treatment was conducted. He also noticed a
decrease of Saccharomyces cerevisiae (5.8 log CFU/mL) when cells were suspended in potassium
phosphate buffer in a 110 mm diameter watch glass treated with PL of total of 3.5 J/cm2 energy
from a flash lamp. A reduction in L. monocytogenes count of 2.4 log CFU/mL was observed after
inoculating 7 log CFU/mL on skinless chicken breast meat after pulsed treatment of 5.4 J/cm2
(Paskeveiciute et al., 2010). Pulsed UV light was effective in reducing the population of Salmonella
enteritidis on the surface of shell eggs. In curds of commercially dry cottage cheese which was
inoculated with Pseudomonas spp., the application of PL with energy of 16 J/cm2 reduced microbial
population by 1.5 log cycles (Dunn et al., 1991). Dunn et al. (1989) had worked on the browning
of potato slices and claimed that by using two to five flashes of light at a fluence of 3 J/cm2, the
browning can be inhibited. The treated slices exhibited less PPO (polyphenol oxidase) activity than
that from the untreated slices. PL can also increase the respiration rate of vegetables. Dunn et al.
(1997) observed that beef steaks treated with 5 J/cm2 PL on both sides exhibited 2-log reductions in
microbial counts upon storage for 3 days at 4°C–5°C. Dunn et al. (1995) reported a 2-log reduction
in Listeria innocua in hot dogs after treatment with PL. Ozer and Demirci (2006) demonstrated
1-log reduction of E. coli O157:H7 and/or L. monocytogenes in salmon fillets after a PL treatment
(5.6 J/cm2 per pulse) for 60 s at 8 cm distance without affecting the quality. Shrimp treated with PL
had better shelf life and remained edible for 7 days, whereas untreated shrimp showed extensive
microbial degradation and lead to discoloration, foul smelling and became inedible (Dunn et al.,
1995). But Dunn et al. (1989) had observed that PL was able to reduce the psychrotroph and coliform
population on the surface of summer flounder fillets and the fillets remained sensorily acceptable
after 15 days of refrigerated storage.
Pollock (2007) focused on the PL destruction of L. monocytogenes on vacuum-packed cold-

smoked salmon, and have shown that combined effect with low-temperature storage (4°C),
PL had the potential to extend the shelf-life without compromising sensory quality. Shuwaish
et al. (2000) found that application of 2–4 pulses of 2.5–5 J/cm 2 had no significant change in
Hunter colour values nor shear force values in pre-packaged catfish fillets. Dunn et al. (1995)
found PL treatment of 30J/cm 2 did not cause any loss of proteins, riboflavin, and ascorbic acid
in frankfurters and riboflavin content in beef, chicken, and fish. Another application of PL is
surface sterilization of food packaging materials and on the surface of packaged products in UV
transparent materials.
18.16 ULtraSOUND prOCESSING
The application of ultrasound in food processing has been stared as another area in non-thermal
approaches, which exploits the preservative effect of the high-intensity sound waves. The pre-
servative effect is by the inactivation of microbes and spoilage enzyme by mechanical actions.
Ultrasonic cavitation when propagated through biological structures produces shear forces, which
causes mechanical cell breakage and allows material transfer from cell into solvents. Cavitation
causes particle size reduction thereby increases the surface area in contact while extracting a com-
pound. The technology find its application in the field of extraction of proteins, lipids and their
functional modifications, emulsification, viscosity improvement, homogenization and improvement
of dispersion stability in liquid foods (Adzahan and Benchamaporn, 2007). Therefore, this technol-
ogy is utilized in the field of processing, preservation and extraction, which makes use of physical
and chemical phenomena that are fundamentally different from conventional extraction, process-
ing or preservation techniques. Advantages include better productivity, yield and selectivity, better
processing time, enhanced quality, reduced chemical and physical hazards, and environmentally
friendly (Chemat et al., 2011).
Ultrasonic characterization of Atlantic mackerel was done by Sigfusson et al. (2001) using
a semi-empirical equation in which fat, water and solids-non-fat content of mackerel tissue was
determined by analysing the temperature dependence of the ultrasonic velocity by different
methods. They observed a good agreement between the fat, water and solids-non-fat content as
determined by ultrasonic velocity and proximate analysis methods. The increase in ultrasonic
attenuation of mackerel with temperature, especially above 20°C, was attributed to protein
denaturation and tissue disruption. Ghaedian et al. (1998) studied the association between the
ultrasonic properties of fish and their composition. The temperature dependence of the ultrasonic
velocity of fish analogues was measured from 5°C to 35°C and it was observed that ultrasonic
velocity increased with solids-non-fat at all temperatures, but had a more complex dependence
on fat content. The ultrasonic velocity was independent of fat at around 15°C, and increased with
fat content at lower temperatures and decreased at higher temperatures. Research highlighted
the potential use of ultrasonic velocity measurements to determine the composition of fish in a
more rapid and non-destructive method. Another area of investigation is the effects of ultrasonic
wave treatment on the extraction yield of acid soluble collagen from sea bass skins (Kim et al.,
2012). A comparison on extraction of collagen with 24 h acid treatment using 0.5 M acetic acid
(1:200 sample/acid, w/v) and an extraction using ultrasonic treatment after the addition of a 0.5 M
acetic acid solution was done and they observed an increase in extraction yield of collagen during
ultrasonic treatment, with noticed higher rate of extraction at higher amplitudes of ultrasonic
treatment. Another possibility is in the line of acid /alkaline extraction assisted by ultrasounds and
it is possible to recover more than 95% of total protein from mackerel by-products. The researches
on application of ultrasound in seafood processing and preservation is minimal, and there is a
future scope for ultrasound in fish processing industries.
18.17 CONCLUSION
Food safety and quality are the main dynamic powers behind today’s consumer acceptance
of a product/technology. This has created an urge towards the introduction and adoption of new
technologies that can enhance the nutritional and sensory qualities of the food. Seafoods, being
a delicacy, need a novel processing method for retaining the freshness and quality. Out of the dif-
ferent non-thermal technologies in vogue, only HPP has commercial application in sea foods. Its
application is varied depending on the end product and end users and in spite of its initial high
capital investment, the benefits of high pressure are recognized. A large number of units have come
up and manufactures bring out desired size and capacity machines in an automated way. PL and
ultrasound processing are in the nascent stage and have yet to be commercially exploited in seafood.
The benefits in the food processing are limited and hence future research will be really slow. Non-
thermal techniques find themselves a niche market segment where unique high-ended products are
placed. A future approach is needed for functional modifications of food macromolecules using
non-thermal techniques.
rEFErENCES
Adzahan, M. N., and Benchamaporn, P. (2007). Potential of non-thermal processing for food preservation in
Southeast Asian Countries. ASEAN Food J. 14(3): 141–152.
Ahn, J., Balasubramaniam, V. M., and Yousef, A. E. (2007). Inactivation kinetics of selected aerobic and
anaerobic bacterial spores by pressure-assisted thermal processing. Int. J. Food Microbiol. 113(3):
321–329.
Alizadeh, E., Chapleau, N., De Lamballerie, M., and Le Bail, A. (2007). Effect of different freezing pro-
cesses on the microstructure of Atlantic salmon (Salmo salar) fillets. Innov. Food Sci. Emerg. 8(4):
493–499.
Amanatidou, A., Schluter, O., Lemkau, K., Gorris, L. G. M., Smid, E. J., and Knorr, D. (2000). Effect of
combined application of high pressure treatment and modified atmospheres on the shelf life of fresh
Atlantic salmon. Innov. Food Sci. Emerg. 1(2): 87–98.
Angsupanich, K., and Ledward, D. A. (1998). High pressure treatment effects on cod (Gadusmorhua) muscle.
J. Food Chem. 63(1): 39–50.
Ashie, I. N. A., and Simpson, B. K. (1996). Application of high hydrostatic pressure to control enzyme related
fresh seafood texture deterioration. Food Res. Int. 29(5–6): 569–575.
Ashie, I. N. A., and Lanier, T. C. (1999). High pressure effects on gelation of surimi and turkey breast muscle
enhanced by microbial transglutaminase. J. Food Sci. 64: 704–708.
Ashie, I. N. A., Simpson, B. K., and Ramaswamy, H. S. (1997). Changes in texture and microstructure of
pressure-treated muscle tissue during chilling storage. J. Muscle Foods. 8: 12–32.
Ashie, I. N. A., Simpson, B. K., and Ramaswamy, H. S. (2007). Changes in texture and microstructure of
pressure-treated fish muscle during chilled storage. J. Muscle Foods. 8(1): 13–32.
Balasubramaniam, V. M., and Farkas, D. (2008). High-pressure food processing. Food Sci. Technol. Int. 14(5):
413–418.
Balasubramanian, V. M., Ting, E. Y., Stuart, C. M., and Robbins, J. A. (2004). Recommended laboratory
practices for conducting high pressure microbial inactivation experiments. Innov. Food Sci. Emerg. 5:
299–306.
Bank, H. L., John, J., Schmehl, M. K., and Dratch, R. J. (1990). Bactericidal effectiveness of modulated UV
light. Appl. Environ. Microbiol. 56: 3888–3889.
Barbosa-Cánovas, G. V., and Bermudez-Aguirre, D. (2010). Other milk preservation technologies: Ultrasound,
irradiation, microwave, radio frequency, ohmic heating, ultraviolet light and bacteriocins. In: Improving
the Safety and Quality of milk, vol. 1, Griffiths, M., Ed. Cambridge, UK: Woodhead.
Bindu, J., Ginson, J., Kamalakanth, C. K., and Gopal, T. K. S. (2015). High pressure treatment of green mussel
Pernaviridis Linnaeus, 1758: Effect on shucking and quality changes in meat during chill storage.
Indian J. Fish. 62(2): 70–76.
Campus, M. (2010). High pressure processing of meat, meat products and seafood. Food Eng. Rev. 2(4):
256–273.
Cando, D., Moreno, H. M., Tovar, C. A., Herranz, B., and Borderias, A. J. (2014). Effect of high pressure and/
or temperature over gelation of isolated hake myofibrils. Food Bioprocess Tech. 7: 3197–3207.
Carlez, A., Veciana-Nogues, T., and Cheftel, J. C. (1995). Changes in colour and myoglobin of minced beef
meat due to high pressure processing. Lebensmittel-Wissenschaft und-Technologie. 28(5): 528–538.
Carpi, G., Gola, S., Maggi, A., Rovere, P., and Buzzoni, M. (1995). Microbial and chemical shelf-life of high
pressure treated salmon cream at refrigeration temperatures. Ind. Converse. 70(4): 386–397.
Cheftel, J. C. (1995). Review: High-pressure, microbial inactivation and food preservation. Food Sci. Technol.
Int. 1: 75–90.
Cheftel, J. C., and Culioli, J. (1997). Effects of high pressure on meat: A review. Meat Sci. 46(3): 211–236.
Chemat, F., Muhammed, Z., and Khan, K. (2011). Applications of ultrasound in food technology: Processing,
preservation and extraction. Ultrason. Sonochem. 18(4): 813–835.
Chéret, R., Chapleau, N., Delbarre-Ladrat, C., Verrez-Bagnis, V., and Lamballerie, M. D. (2005). Effects of high
pressure on texture and microstructure of sea bass (Dicentrarchus labrax L.) fillets. J. Food Sci. 70(8): 477–483.
Chéret, R., Delbarre-Ladrat, C., Lamballerie-Anton, M., and Verrez-Bagnis, V. (2007). Calpain and cathepsin
activities in post mortem fish and meat muscles. Food Chem. 101: 1474–1479.
Considine, K. M., Kelly, A. L., Fitzgerals, G. F., Hill, C., and Sleator, R. D. (2008). High pressure processing
effects on microbial food safety and food quality. FEMS Microbiol. Letters. 281(1): 1–9.
Cruz-Romero, M., Kelly, A. L., and Kerry, J. P. (2007). Effects of high-pressure and heat treatments on physical
and biochemical characteristics of oysters (Crassostreagigas). Innov. Food Sci. Emerg. 8(1): 30–38.
Dunn, J., Bushnell, A., Ott, T., and Clark, W. (1997). Pulsed white light food processing. Cereal Food. World.
42: 510–515.
Dunn, J., Ott, T., and Clark, W. (1995). Pulsed light treatment of food and packaging. Food Technologist.
49(9): 95–98.
Dunn, J. E., Clark, W. R., and Asmus, J. F. (1989). Methods for Preservation of Foodstuffs. Maxwell
Laboratories Inc., San Diego, CA. US Patent 4871559.
Dunn, J. E., Clark, W. R., and Asmus, J. F. (1991). Methods for Preservation of Foodstuffs. Maxwell
Laboratories Inc., San Diego, CA. US Patent 5034235.
Eisenbrand, G. (2005). Safety assessment of high pressure treated foods. Mol. Nutr. Food Res. 49(12):
1168–1174.
Erkan, N., Uretener, G., and Alpas, H. (2010). Effect of high pressure (HP) on the quality and shelf life of red
mullet (Mullussurmelutus). Innov. Food Sci. Emerg. 11(2): 259–264.
Errol, V. R. (2007). High Hydrostatic Pressure Processing of Seafood. Avure Technologies, USA.
Evans, G., and Cox, D. N. (2006). Australian consumers’ antecedents of attitudes towards foods produced by
novel technologies. Br. Food J. 108(11): 916–930.
Farkas, J. (2007). Physical methods of food preservation. In: Food Microbiology: Fundamentals and Frontiers,
pp. 685–705. Doyle, P., Beuchat, L. R., and Montville, T. J., Eds. Washington, DC: American Society
for Microbiology Press.
Fernandez-Martin, F., Perez-Mateos, M., and Montero, P. (1998). Pressure/heat combinations on blue whiting
(Micromesitiuspoutassou) thermal and mechanical behaviour. J. Agric. Food Chem. 46(8): 3257–3264.
Gekko, K., and Koga, S. (1983). The effect of pressure on thermal stability and in vitro 16 fibril formation
collagen. Agric. Biol. Chem. 47: 1027–1033.
Ghaedian, R., Coupland, J. N., Decker, E. A., and McClements, D. J. (1998). Ultrasonic determination of fish
composition. J. Food Eng. 35: 323–337.
Ginson, J., Kamalakanth, C. K., Bindu, J., Venkateswarlu, R., Das, S., Chauhan, O. P., and Srinivasa Gopal,
T. K. (2013). Changes in K Value, microbiological and sensory acceptability of high pressure processed
Indian white prawn (Fenneropenaeusindicus). Food Bioprocess Tech. 6(5): 1175–1180.
Gómez-Guillén, M. C., Giménez, B., and Montero, P. (2005). Extraction of gelatin from fish skins by high
pressure treatment. Food Hydrocolloids. 19(5): 923–928.
Gómez-López, V. M., Ragaert, P., Debevere, J., and Devlieghere, F. (2007). Pulsed light for food
decontamination: A review. Trends Food Sci. Technol. 18(9): 464–473.
Gould, G. W. (1977). Recent advances in the understanding of resistance and dormancy in bacterial spores.
J. Appl. Bacteriol. 42: 297–309.
Gould, G. W. (2000). Preservation: Past, present and future. Brit. Med. Bull. 56: 84–96.
Gram, L., and Huss, H. H. (2000). Flesh and processed fish and shellfish. In: The Microbiological Safety
and Quality of Food. pp. 472–506. Lund, B. M., Baird-Parker, T. C., and Gould, G. W., Eds., Vol. 1.
Maryland, MD: Aspen Publishers.
Grauwet, T., Rauh, C., Van der Plancken, I., Vervoort, L., Hendrickx, M., and Delgado, A. (2012). Potential
and limitations of methods for temperature uniformity mapping in high pressure thermal processing.
Trends Food Sci. Technol. 23(2): 97–110.
Gudbjornsdottir, B., Jonsson, A., Hafsteinsson, H., and Heinz, V. (2010). Effect of high-pressure processing
on Listeria spp. and on textural and micro structural properties of cold smoked salmon. LWT – Food
Sci. Tech. 43: 366–374.
Hayakawa, I., Linko, Y., and Linko, P. (1996). Novel mechanical treatments of biomaterials. Lebenson Wiss
Technol. 29(5–6): 395–403.
Hayashi, R. (2002). High pressure in bioscience and biotechnology: Pure science encompassed in pursuit of
value. Biochim. Biophys. Acta – Protein Struct. Molec. Enzym. 1595(1–2): 397–399.
Hayashi, R., Kawamura, Y., Nakasa, T., and Okinaka, O. (1989). Application of high-pressure to food pro-
cessing; pressurization of egg white and yolk, and properties of gels formed. Agric. Biol. Chem. 53:
2935–2939.
He, H., Adams, R. M., Farkas, D. F., and Morrissey, M. T. (2002). Use of high pressure processing for oyster
shucking and shelf-life extension. J. Food Sci. 67: 640–645.
Heinz, I. V., and Buckow, R. (2010). Food preservation by high pressure. J. Verbraucherschutz
Lebensmittelsicherheit. 5: 73–78.
Heinz, V., and Knorr, D. (1998). High pressure germination and inactivation kinetics of bacterial spores. In:
High Pressure Food Science, Bioscience and Chemistry. pp. 435–441. Isaacs, N. S., Ed. Cambridge,
UK: The Royal Society of Chemistry.
Heinz, V., and Knorr, D. (2002). Effects of high pressure on spores. In: Ultra High Pressure Treatment of Foods.
pp. 77–114. Hendrickx, M. E. G., and Knorr, D., Eds. New York, Kluwer Academic/plenium publishers.
Heremans, K. (1982). High pressure effects on proteins and other biomolecules. Annu. Rev. Biophys. Bioeng.
11: 1–21.
Heremans, K. (1995). High pressure effects on biomolecules. In: High Pressure Processing of Foods.
pp. 81–97. Ledward, D. A., Johnston, D. E., Earnshaw, R. G, and Hasting, A. P. M., Eds. Leicestershire,
UK: Nottingham University Press.
Herranz, B., Tovar, C. A., Borderiasa, A. J., and Moreno, H. M. (2013). Effect of high-pressure and/or microbial
transglutaminase on physicochemical, rheological and microstructural properties of flying fish surimi.
Innov. Food Sci. Emerg. 20: 24–33.
Hoover, D. G., Metrick, C., Papineau, A. M., Farkas, D. F., and Knorr, D. (1989). Biological effects of high
hydrostatic pressure on food microorganisms. Food Technol. 43(3): 99–107.
Hultin, H. O. (1994). Oxidation of lipids in seafoods. In: Seafoods: Chemistry, Processing Technology and
Quality. pp. 49–74. Shahidi, F., and Botta, J. F., Eds. Glasgow, UK: Blackie Academic and Professional.
Hurtado, J. L., Montero, P., and Borderías, A.J. (2000). Extension of shelf life of chilled hake
(Merlucciuscapensis) by high pressure. Food Sci. Technol. Int. 6(3): 243–249.
Hurtado, J., Montero, P., Borderías, J., and Solas, M. (2001). High-pressure/temperature treatment effect on
the characteristics of octopus (Octopus vulgaris) arm muscle. Eur. Food Res. Technol. 213(1): 22–29.
Jimenez-Colmenero, F. (2002). Muscle protein gelation by combined use of high pressure/temperature. Trends
Food Sci. Technol. 13(1): 22–30.
Kamalakanth, C. K., Ginson, J., Bindu, J., Venkateswarlu, R., Das, S., Chauhan, O. P., and Gopal, T. K.
S. (2011). Effect of high pressure on K-value, microbial and sensory characteristics of yellowfin tuna
(Thunnusalbacares) chunks in EVOH films during chill storage. Innov. Food Sci. Emerg. Technol.
12(4): 451–455.
Kaur, B., Kaushik, N., Rao, P. S., and Chauhan, O. P. (2013). Effect of high-pressure processing on physical,
biochemical, and microbiological characteristics of black tiger shrimp (Penaeus monodon). Food
Bioprocess Tech. 6(6): 1390–1400.
Kennish, J. M., and Kramer, D. E. (1986). A review of high pressure chromatographic methods for measuring
nucleotide degradation in fish muscle. In: Seafood Quality Determination. pp. 209–219. Proceedings
of the International Symposium Coordinated by the University of Alaska. Sea Grant College Program,
Anchorage, Alaska, U. S. A., Kramer, D. E., and Liston, J., Eds. Amsterdam, the Netherlands: Elsevier
Science Publishers V.
Kim, H. K., Kim, Y. H., Kim, Y. J., Park, J. H., and Lee, N. H. (2012). Effects of ultrasonic treatment on
collagen extraction from skins of the sea bass Lateolabrax japonicas. Fish. Sci. 78: 485–490.
Knoerzer, K., Juliano, P., Gladman, S., and Versteeg, C. (2007). A computational model for temperature and
sterility distributions in a pilot-scale high pressure high-temperature process. AlChE J. 53: 2996–3010.
Knorr, D. (1999). Novel approaches in food-processing technology: New technologies for preserving foods
and modifying function. Curr. Opin. Biotechnol. 10(5): 485–491.
Knorr, D. (2002). High pressure processing for preservation, modification and transformation of foods. High
Pressure Res. 22: 595–599.
Knorr, D. (2003). Impact of non-thermal processing on plant metabolites. J Food Eng. 56(2–3): 131–134.
Knorr, D., Reineke, K., Mathys, A., Heinz, V., and Buckow, R. (2011). High-pressure-induced effects on
bacterial spores, vegetative microorganisms, and enzymes. In: Food Engineering Interfaces. pp.
325–340. Food Engineering Series, Aguilera, J. M., Simpson, R., Welti-Chanes, J., Bermudez-Aguirre,
D., and Barbosa-Canovas, G., Eds. New York: Springer.
Ko, W. C. (1996). Effect of high pressure on gelation of meat paste and inactivation of actomyosin Ca-ATPase
prepared from Milkfish. Fisheries Sci. 62: 101–104.
Koo, J., Jahncke, M. L., Reno, P. W., Hu, X., and Mallikarjunan, P. (2006). Inactivation of Vibrio
parahaemolyticusand Vibrio vulnificusin phosphate-buffered saline and in inoculated whole oysters by
high-pressure processing. J. Food Protect. 69(3): 596–601.
Koutchma, T., Keller, S., Chirtel, S., and Parisi, B. (2004). Ultraviolet disinfection of juice products in laminar
and turbulent flow reactors. Innov. Food Sci. Emerg. 5: 179–189.
Krishnamurthy, K., Tewari, J., Irudayaraj, J., and Demirci, A. (2010). Microscopic and spectroscopic
evaluation of inactivation of Staphylococcus aureus by pulsed UV light and infrared heating. Food
Bioprocess Tech. 3: 93–104.
Kunnath, S., Panda, S. K., Jaganath, B., and Gudipati, V. (2015). Effect of high pressure processing on textural
and microbiological quality of pink perch (Nemipterusjaponicus) sausage during chilled storage. High
Pressure Res. 35(4): 419–429.
Kwiatkowska, A., Jankowska, B., and Korzeniowski, W. (2001). Changes in solubility of the bovine semitendinosus
muscle collagen under the influence of high pressure. Pol. J. Food Nutr. Sci. 10/51(4): 35–39.
Lakshmanan, R., Miskin, D., and Piggott, J. R. (2005). Quality of vacuum packed cold-smoked salmon during
refrigerated storage as affected by high-pressure processing. J. Sci. Food Agr. 85(4): 655–661.
Lakshmanan, R., Parkinson, J. A., and Piggott, J. R. (2007). High-pressure processing and water-holding
capacity of fresh and cold-smoked salmon (Salmo salar). LWT – Food Sci. Tech. 40(3): 544–551.
Lau, M. H., and Turek, E. J. (2007). Determination of quality difference in low-acid foods sterilized by high
pressure versus retorting. In: High Pressure Processing of Foods. pp. 195–217. Doona, J., and Feeherry,
F. E., Eds. Blackwell Publishing and IFT Press, USA.
Le Bail, A., Chevalier, D., Mussa, D. M., and Ghoul, M. (2002). High pressure freezing and thawing of foods:
A review. Int. J. Refrig. 25(5): 504–513.
Leistner, L. (1995). Principles and application of Hurdle Technology. In: New Methods of Food Preservation.
pp. 1–20. Gould, G. W., Ed. Boston, MA: Springer.
Linton, M., McClements, J. M. J., and Patterson, M. F. (2003). Changes in the microbiological quality of
shellfish, brought about by treatment with high hydrostatic pressure. Int. J. Food Sci. Tech. 38: 713–727.
Lopez-Caballero, M. E., Perez-Mateos, M., Bonderıas, A. J., and Montero, P. (2000a). Extension of shelf life
of prawns (Penaeusjaponicus) by vacuum packaging and high-pressure treatment. J. Food Protect. 63:
1381–1388.
Lopez-Caballero, M. E., Perez-Mateos, M., Borderias, A. J., and Montero, P. (2000b). Oyster preservation by
high pressure treatment. J. Food Protect. 63: 196–201.
Lopez-Malo, A., Palou, E., Barbosa-Canovas, G. V., Swanson, B. G., and Welti-Chanes, J. (2000). Minimally
processed foods and high hydrostatic pressure. In: Advances in Food Engineering. pp. 267–286.
Lozano, J., Anon, M. C., Parada-Arias, E., and Barbosa-Canovas, G. V., Eds. Lancaster, PA: Technomic
Publishing Co.
Margosch, D., Ehrmann, M. A., Ganzle, M. G., and Vogel, R. E. (2004). Comparison of pressure and heat
resistance of Clostridium botulinum and other endospores in mashed carrots. J. Food Prot. 67: 2530–2537.
Margosch, D., Ehrmann, M.A., Buckow, R., Heinz, V., Vogel, R. F., and Gänzle, M.G. (2006). High-Pressure-
Mediated Survival of Clostridium botulinum and Bacillus amyloliquefaciens Endospores at High
Temperature. Appl. Environ. Microbiol., 72(5): 3476–3481.
Masson, P. (1992). Pressure denaturation of proteins. In: High Pressure and Biotechnology. pp. 89–99.
Balny, R., Hayashi, K., Heremans, P., and Masson, P., Eds. Vol. 224. Montrouge, France: Colloque
INSERM/John Libbery Eurotext Ltd.
Master, A. M., Stegeman, D., Kals, J., and Bartels, P. V. (2000). Effects of high pressure on colour and texture
of fish. High Pressure Res. 19: 109–115.
Mathys, A. (2008). Inactivation mechanism of zero geo bacillus and bacillus spores during high pressure
thermal sterilization, Thesis (PhD). Technische Universitat. Berlin.
Matser, A. M., Krebbers, B., Van den Berg, R. W., and Bartels, P. V. (2004). Advantages of high pressure
sterilization on quality of food products. Trends Food Sci. Technol. 15(2): 79–85.
Messens, W., Van Camp, J., and Huygebaert, A. (1997). The use of high pressure to modify the functionality
of food proteins. Trends Food Sci. Technol. 8: 107–112.
Montero, P., Gomez-Estaca, J., and Gomez-Guillen, M. C. (2007). Influence of salt, smoke and high pressure on
Listeria monocytogenes and spoilage microflora in cold smoked dolphinfish. J. Food Protect. 70: 399–404.
Montero, P., Pérez-Mateos, M., and Solas, T. (1997). Comparison of different gelation methods using washed
sardine (Sardinapilchardus) mince: Effects of temperature and pressure. J. Agric. Food Chem. 45:
4612–4618.
Mújica-Paz, H., Valdez-Fragoso, A., Tonello-Samson, C., Welti-Chanes, J., and Torres, J. A. (2011). High-
pressure processing technologies for the pasteurization and sterilization of foods. Food Bioprocess
Technol. 4: 969–985.
Murchie, L. W., Cruz-Romero, M., Kerry, J. P., Linton, M., Patterson, M. F., Smiddy, M., and Kelly, A. L.
(2005). High pressure processing of shellfish: A review of microbiological and other quality aspects.
Innov. Food Sci. Emerg. Tec. 6(3): 257–270.
Nagashima, Y., Ebina, H., Tanaka, M., and Taguchi, T. (1993). Effect of high hydrostatic pressure on the ther-
mal gelation of squid mantle meat. Food Res. Int. 26(2): 119–123.
Narwankar, S. P., Flimlin, G. E., Schaffner, D. W., Tepper, B. J., and Karwe, M. V. (2011). Microbial safety
and consumer acceptability of highpressure processed hard clams (Mercenariamercenaria). J. Food
Sci. 76(6): M375–M380.
National Center for Food Safety and Technology. (2009). NCFST receives regulatory acceptance of novel food
sterilization process. Press release, February 27, Summit-Argo, IL.
Nuray, E., Uretener, G., and Alpas, H. (2010). Effect of high pressure (HP) on the quality and shelf life of red
mullet (Mullussurmelutus). Innovt. Food Sci. Emerg. Tech. 11(2): 259–264.
Offer, G., and Trinick, J. (1983). On the mechanism of water holding in meat: The swelling and shrinking of
myofibrils. Meat Sci. 8(4): 245–281.
Ohshima, T., Ushio, H., and Koizumi, C. (1993). High-pressure processing of fish and fish products. Trends
Food Sci. Tech. 4(11): 370–375.
Olsen, N. V., Grunert, K. G., and Soiine, A. M. (2010). Consumer acceptance of high pressure processing and
pulsed electric field: A review. Trends Food Sci. Technol. 21: 464–472.
Ozer, N. P., and Demirci, A. (2006). Inactivation of Escherichia coli O157: H7 and Listeria monocyto-
genes inoculated on raw salmon fillets by pulsed UV light treatment. Int. J. Food Sci. Tech. 41:
354–360.
Food Processing. pp. 279–306. Sun, D.-W., Ed. London, UK: Academic Press.
Paskeveiciute, E., Buchovec, I., and Luksiene, Z. (2010). High power pulsed light for decontamination of
chicken from food pathogens: A study on antimicrobial efficiency and organoleptic properties. J. Food
Safety. 31: 61–68.
Patterson, M. F. (2005). Microbiology of pressure treated foods. J. Appl. Microbiol. 98(6): 1400–1409.
Patterson, M. F., Linton, M., McCiements, J. M. J., Farmer, L. J., Johnston, D. E., and Dynes, C. (2003). Effect
of high hydrostatic pressure on the microbiological, chemical and sensory quality of fish. IFT Annual
Meeting-2003 Chicago.
Pollock, A. M. (2007). M.Sc. thesis. Department of Food Science and Agricultural Chemistry, Macdonald
Campus of McGill University. Canada.
Potasman, I., Paz, A., and Odeh, M. (2002). Infectious outbreaks associated with bivalve shellfish consumption:
A worldwide perspective. Clin. Infect. Dis. 35(8): 921–928.
Ramirez, R., Saraiva, J., Perez Lamela, C., and Torres, J. A. (2009). Reaction kinetics analysis of chemical
changes in pressure-assisted thermal processing. Food Eng. Rev. 1(1): 16–30.
Ramirez-Suarez, J. C., and Morrissey, M. T. (2006). Effect of high pressure processing (HPP) on shelf life of
albacore tuna (Thunnusalalunga) minced muscle. Innov. Food Sci. Emerg. 7(1–2): 19–27.
Rastogi, N. K., Raghavarao, K. S. M. S., Balasubramaniam, V. M., Niranjan, K., and Knorr, D. (2007).
Opportunities and challenges in high pressure processing of foods. Crit. Rev. Food Sci. Nutr. 47: 69–112.
Reineka, K. (2012). Mechanics of Baccillus spore germination and inactivation during high pressure
processing. Thesis (Ph. D). Technische Universitat. Berlin.
Reineke, K., Mathys, A., Heinz, V., and Knorr, D. (2013). Mechanism of endospore inactivation under high
pressure. Trends Microbiol. 21(6): 296–304.
Richards, G. P. (1988). Microbial purification of shellfish: A review of depuration and relaying. J. Food Prot.
51: 218–251.
Rode, T. M., and Hovda, M. B. (2016). High pressure processing extend the shelf life of fresh salmon, cod and
mackerel. Food Control. 70: 242–248.
Rouille, J., Lebail, A., Ramaswamy, H. S., and Leclerc, L. (2002). High pressure thawing of fish and shellfish.
J. Food Eng. 53(1): 83–88.
Sale, A. J., Gould, G. W., and Hamilton, W. A. (1970). Inactivation of bacterial spores by hydrostatic pressure.
J. Gen. Microbiol. 60: 323–334.
Sareevoravikul, R., Simpson, B. K., and Ramaswamy, H. S. (1996a). Comparative properties of bluefish gels
formulated by high pressure and heat. J. Aquatic Food Prod. Technol. 5: 65–79.
Sequeira-Munoz, A., Chevalier, D., LeBail, A., Ramaswamy, H. S., and Simpson, B. K. (2006). Physicochemical
changes induced in carp (Cyprinuscarpio) fillets by high pressure processing at low temperature. Innov.
Food Sci. Emerg. 7(1–2): 13–18.
Setlow, B., Cowan, A. E., and Setlow, P. (2003). Germination of spores of Bacillus subtilis with dodecylamine.
J. Appl. Mic. 95: 637–648.
Sevenich, R., Rauh, C., and Knorr, D. (2016). A scientific and interdisciplinary approach for high pressure
processing as a future toolbox for safe and high quality products: A review. Innov. Food Sci. Emerg.
38: 65–75.
Shimidzu, N., Goto, M., and Miki, W. (1996). Carotenoids as singlet oxygen quenchers in marine 736
organisms. Fisheries Sci. 62(1): 134–137.
Shuwaish, A., Figueroa, J. E., and Silva, J. L. (2000). Pulsed-light treated pre-packaged catfish fillets. IFT
Annual Meeting, Dal las, USA, 10–14 June, 2000.
Sigfusson, H., Decker, E. A., and McClements, D. J. (2001). Ultrasonic characterization of Atlanticmackerel
(Scomberscombrus). Food Res. Int. 34: 15–23.
Smelt, J. P. M. (1998). Recent advances in the microbiology of high pressure processing. Trends Food Sci.
Technol. 9: 152–158.
Sudheer, K. P., Griet, K., Marc, H., and Ann, V. L. (2011). Effect of HPP on in-vitro bio-accessibility
of lycopene from tomato. In: Compendium of Emerging Technologies in Food Processing for
Ensuring Food Safety and Quality.Abstract No. O-II-11 14-15, October, 2011, TNAU. Foodxplore’11,
Coimbatore.
Sun, X. D., and Holley, R. A. (2010). High hydrostatic pressure effects on the texture of meat and meat
products. J. Food Sci. 75: R17–R23.
Suzuki, A. (2002). High pressure-processed foods in Japan and the world. In: Progress in Biotechnology.
pp. 365–374. Rikimaru, H., Ed. New York: Elsevier.
Suzuki, A., Watanabe, M., Ikeuchi, Y., Saito, M., and Takahashi, K. (1993). Effects of high-pressure treatments
on the ultrastructure and thermal behaviour of beef intramuscular collagen. Meat Sci. 35: 17–25.
Takeshita, K., Shibato, J., Sameshima, T., Fukunaga, S., Isobe, S., Arihara, K., and Itoh, M. (2003). Damage
of yeast cells induced by pulsed light irradiation. Int. J. Food Microbiol. 85: 151–158.
Tanaka, M., Zhuo, X. Y., Nagashima, Y., and Taguchi, T. (1991). Effect of high pressure on the lipid oxydation
in sardine meat. Nippon Suisan Gakk. 57: 957–963.
Terova, G., Preziosa, E., Marelli, S., Gornati, R., Bernardini, G., and Saroglia, M., (2011). Applying
transcriptomics to better understand the molecular mechanisms underlying fish filet quality. Food
Chem. 124(3): 1268–1276.
Ting, E. (2007). Compression heating and temperature control in high pressure processing. In: High Pressure
Processing of Foods. pp. 227–234. Doona, C. J., and Feeherry, F. E., Eds. IFT Press and Blackwell
Publishing, USA.
Toepfl, S., Mathys, A., Heinz, V., and Knorr, D. (2006). Review: Potential of high hydrostatic pressure and
pulsed electric fields for energy efficient and environmentally friendly food processing. Food Rev. Int.
22: 405–423.
Torres, B., Tiwari, B. K., Patras, A., Cullen, P. J., Brunton, N., and O’Donnell, C. P. (2011). Stability of
anthocyanins and ascorbic acid of high pressure processed blood orange juice during storage. Innov.
Food Sci. Emerg. 12: 93–97.
Torres, J. A., and Velazquez, G. (2005). Commercial opportunities and research challenges in the high pressure
processing of foods. J. Food Eng. 67: 95–112.
Truong, B. Q., Buckow, R., Nguyen, M. H., and Stathopoulos, C. E. (2016). High pressure processing of
barramundi (Latescalcarifer) muscle before freezing: The effects on selected physicochemical
properties during frozen storage. J. Food Eng. 169: 72–78.
Uesugi, A. R., and Moraru, C. I. (2009). Reduction of Listeria on ready-to-eat sausages after exposure to a
combination of pulsed light and nisin. J. Food Prot. 72(2): 347–353.
Uresti, R. M., Velazquez, G., Ramirez, J. A., Vazquez, M., and Torres, J. A. (2004). Effect of high-pressure
treatments on mechanical and functional properties of restructured products from arrowtooth flounder
(Atheresthesstomias). J. Sci. Food Agric. 84(13): 1741–1749.
Uresti, R. M., Velazquez, G., Vazquez, M., Ramirez, J. A., and Torres, J. A. (2005). Effect of sugars and
polyols on the functional and mechanical properties of pressure-treated arrowtooth flounder
(Atheresthesstomias) proteins. Food Hydrocolloid. 19(6): 964–973.
Uresti, R. M., Velazquez, G., Vazquez, M., Ramirez, J. A., and Torres, J. A. (2006). Effects of combining
microbial transglutaminase and high pressure processing treatments on the mechanical properties of
heat-induced gels prepared from arrow tooth flounder (Atheresthesstomias). Food Chem. 94(2): 202–209.
Vázquez, M., Torres, J. A., Gallardo, J. M., Saraiva, J., and Aubourg, S. P. (2013). Lipid hydrolysis and
oxidation development in frozen mackerel (Scomberscombrus): Effect of a high hydrostatic pressure
pre-treatment. Innov. Food Sci. Emerg. 18: 24–30.
Wada, S., and Ogawa, Y. (1996). High pressure effects on fish lipid degradation: Myoglobin change and water
holding capacity. Progr. Biotechnol. 13: 351–356.
Wan, J., Coventry, J., Swiergon, P., Sanguansri, P., and Versteeg, C. (2009). Advances in innovative processing
technologies for microbial inactivation and enhancement of food safety—pulsed electric field and low-
temperature plasma. Trends Food Sci. Technol. 20(9): 414–424.
Wimalaratne, S. K., and Farid, M. M. (2008). Pressure assisted thermal sterilization. Food Bioprod. Process.
86: 312–316.
Xiong, Y. L. (1997). Structure-Function relationships of muscle proteins. In: Food Proteins and Their
Applications. pp. 341–392. Damodaran, S., and Paraf, A., Eds. New York: Marcel Dekker.
Yagiz, Y., Kristinsson, H. G., Balaban, M. O., and Marshall, M. R. (2007). Effect of high pressure treatment
on the quality of rainbow trout (Oncorhynchus mykiss) and Mahi Mahi (Colyphaenahippurus). J. Food
Sci. 72(9): 509–515.
Yagiz, Y., Kristinsson, H. G., Balaban, M. O., Welt, B. A., Ralat, M., and Marshal, M. R. (2009). Effect of
high pressure processing and cooking treatment on the quality of Atlantic salmon. Food Chem. 116:
828–835.
Yoshioka, K., and Yamamoto, T. (1998). Changes of ultrastructure and the physical properties of carp muscle
by high pressurization. Fisheries Sci. 64(1): 89–94.
Zare, Z. (2004). High pressure processing of fresh tuna fish and its effects on Shelf Life. MSc Thesis. McGill
University, Montreal, Quebec, Canada, pp. 45–76.
ChaptEr 19
packaging requirements for

Non-thermal processed Foods
Poonam Mishra and O. P. Chauhan
CONtENtS
19.1 Introduction ......................................................................................................................... 395

19.2 High-Pressure Processing ................................................................................................... 396
19.2.1 Packaging Requirement for HPP .......................................................................... 397
19.3 Irradiation ........................................................................................................................... 401
19.3.1 Packaging for Irradiation ......................................................................................403
19.4 Pulsed Electric Field ...........................................................................................................406
19.4.1 Packaging Requirement for Pulse Electric Field ..................................................407
19.5 Pulsed Light ........................................................................................................................407
19.5.1 Packaging Requirement for Pulsed Light Treatment ............................................408
19.6 Conclusion...........................................................................................................................409
References ...................................................................................................................................... 410
19.1 INtrODUCtION
Thermal processing of food is the most widely used method for preservation of food. There
are several methods of thermal processing, of which pasteurization and sterilization are the most
commonly used methods. Pasteurization inactivates pathogenic vegetative microorganisms; however,
sterilization kills the entire microorganism including spores in the food sample. Traditional thermal
technology and equipment are well developed but preservation by thermal treatments degrades the
quality of foods in terms of colour, flavour, texture, and nutrients (Chen et al., 2015).
Nowadays, consumers are becoming more health conscious and demand convenient,
high-quality, and minimally processed foods with significant retention of nutrients. Non-thermal
processing is a better alternative to fulfil these demands over conventional thermal techniques
(Morris et al., 2006). Non-thermal techniques include high-pressure processing (HPP), pulsed elec-
tric field (PEF), irradiation, pulsed light (PL), disinfection, and electron beam irradiation (Muredzi,
2012; Vanderroost et al., 2014). The mechanisms of preservation of non-thermal techniques are
comparatively different than those of thermal techniques (Kumar and Han, 2012). The success
of extending the shelf life of non-thermally processed foods greatly depends on packaging and
post-packaging conditions of food. Proper selection of packaging material depends on its mechanical
property, tensile property, barrier property, targeted length of shelf life, and the cost. Exposure
of different non-thermal processing conditions may alter the chemical/physical properties of the
395
packaging material and this alteration may influence the shelf life of the food during storage (Ozen
and Floras, 2001). HPP, irradiation, and PL technology can only be applied to packaged food; thus,
the interaction of food with packaging material during processing should be properly addressed.
The preservation of food with HPP is a promising technology for preserving the food. In HPP,
solid or liquid foods are exposed to high pressure (300–700 MPa) for a short period of time for about
10–15 min to reduce the microbial load and inactivate the enzyme. There are two general scientific
principles which explain the relevance of HPP in food processing. The first one is Le Chatelier’s
principle, which is applicable to all physical processes and states that when a system at equilibrium
is disturbed then the system response in a such a way that tends to minimize the disturbance (Norton
et al., 2008). So, any phenomena, i.e., phase transition, change in molecular configuration, and
chemical reactions which are accompanied by a decrease in volume, will be enhanced by pressure
(Tao et al., 2007). Second, the isostatic rule states that if the sample is in intimate contact with the
pressure the pressure will instantaneously and uniformly be transmitted through the sample (Norton
et al., 2008). Pressure transmitted in a uniform and quasi-instantaneous manner, so in contrast to ther-
mal processing time necessary for pressure processing, is independent of the sample size. Physical
compression of the product under high pressure increases the temperature of the product only during
the treatment and this unique property of HPP reduces the severity of thermal effects encountered
with conventional thermal preservation techniques. The heat generated during compression raises
the temperature of product 3°C for every 100 MPa, a change reversed during decompression (Morris
et al., 2006). Other advantages of HPP include that it only disrupts hydrogen bonds and hydrophobic
and electrostatic interactions but has no effect on covalent bonds and, thus, has little effect on chemi-
cal constituents of foods, which is very much desirable to maintain the quality of food (Panadrangi
and Balasubramaniam, 2005). In HPP process, the food product to be treated is placed in a pres-
sure vessel (capable of sustaining high pressure) and submerged in a pressure-transmitting liquid
medium. Water, castor oil, silicone oil, ethanol, or glycol can be used as the pressure-transmitting
medium. The pressure-transmitting liquid is compressed up to a desired pressure and once the pres-
sure is attained the pump or piston is stopped, the valves are closed, and pressure is maintained for a
predetermined time; after that, the system is depressurized (Hogan et al., 2005). Microbial inactiva-
tion by HPP is extensively studied and reviewed by several scientists and has been concluded to be
the result of a combination of factors. HPP causes deprotonation of charged group and disruption
of salt bridges and hydrophobic bonds in cell membranes which causes change in cell morphology,
denaturation of protein and key enzyme, and inhibition of genetic mechanism. These conformational
changes cause the death of microorganisms (Linton and Patterson, 2000; FDA/CFSAN, 2005). Most
vegetative cells can be inactivated at relatively low pressure, i.e., 200–400 MPa (Cheftel, 1995);
however, bacterial spores are more resistant and only very high exposure of pressure (>800 MPa)
can kill bacterial spores. HPP can be used for both pasteurization and sterilization of food products.
In high-pressure pasteurization the food products are exposed with moderate pressures (between
400 and 600 MPa) and at 60°C temperature. High-pressure pasteurization inactivates pathogenic
vegetative microorganism effectively in foods with nutrient degradation (Cheftel, 1995; Farkas and
Hoover, 2000; Raghubeer et al., 2000). The products pasteurized by HPP require refrigeration during
storage and transportation (Pandrangi and Balasubramanian, 2005). For sterilization, exposure of
about 800 MPa at 120°C is required to make the shelf-stable food free from pathogenic bacterial
spores (Meyer et al., 2000; Balasubramanian and Balasubramaniam, 2003). An alternative to treat-
ments combining of pressure and heat for killing of bacterial spores is first to allow the bacterial
spores to germinate and then expose with HPP to kill the bacterial spores. Yeast and moulds are
generally more susceptible to pressure than to bacterial spores so they can be easily inactivated at
PACKAGinG reQuireMents For non-tHerMAL ProCesseD FooDs 397
relatively low pressure. Yeast is simply single-celled fungi and treatment of less than 400 MPa for a
few minutes is sufficient to inactive most of the yeasts. At about 100 MPa, the nuclear membrane of
yeast is affected and at more than 400–600 MPa, alteration in mitochondria and cytoplasm occurs,
which causes the death of the yeast (Smelt, 1998).
HPP can be applied for both liquid and solid foods. Acidic foods are normally good applicants
for HPP; however, HPP is not very successful for low-acidic foods (Clark, 2006). HPP processing
can cause shift in pH of foods and affects enzymatic reactions by changing the structure of
enzymes or substrate (Hernandez-Andres et al., 2005). HPP-processed products like fruits, jellies,
jam, fruit juices, tomato ketchup, ham, chicken, strips, etc. are commercially available in America
(Caner et al., 2004; Devlieghere et al., 2004).
19.2.1 packaging requirement for hpp
As described earlier, food products should be packed in flexible packaging material prior to HP
processing in a batch system, to overcome probable volume reduction in the food in the package and
also to control the collapse of the head space (Ozen and Floros, 2001). Packaging material for HPP
should restored from the deformation due to pressure to its original shape without any significant
changes in physical or chemical properties (Han, 2007). Packaging material widely used for HPP
is given in Table 19.1 (Min et al., 2017). HPP processing requires airtight packages that can resist
the change in volume due to the compressibility of the sample (Hugas et al., 2002), as decrease in
table 19.1 packaging Materials for Foods processed by pulsed Electric Fields, high pressure and
Irradiation
processing packaging Method and

Method Food packaging Material Storage temperature Sources
PeF orange 200 mL thermoformed plastic thermo-forming and sealing Qiu et al.
juice, a container, glass bottle using an aseptic packaging (1998)
protein- machine, sterilized by heat
fortified and H2o2, storage at 4°C
food
beverage
Cranberry Glass vials sealed with small storage at 4°C and Jin and
juice head space 20°C–24°C Zhang
(2002)
Apple juice Materials: base material, HiPs/ thermoforming and evrendilek
apple PVDC/LDPe (Allista Plastic thermo-sealing using and and Zhang
cider, Packaging Co., Muncie, in); lid aseptic packaging (2003);
orange material, nylon/Ai/LDPe machine, sterilized by heat yeom et al.
juice, (rollprint, Addison, iL). and H2o2. storage (2000);
cranberry thermoforming, size of temperature: apple juice, evrendilek
juice, thermoformed plastic apple cider, cranberry et al. (2001)
chocolate container: apple juice and juice, chocolate milk, 4°C,
milk cider, 180 mL; orange juice, 22°C and 37°C; orange
cranberry juice, chocolate juice, cranberry juice,
milk, 200 mL chocolate milk, 4°C and
22°C
orange 500 mL glass, Pet, LDPe, Packed in a glove box, Ayhan et al.
juice HDPe bottles with 28 mm PP sanitized by H2o2 and uV, (2001)
caps (glass, Pet, HDPe- storage at 4°C and 22°C
General bottles supply Co.,
Los Angeles, CA; LDPe-
Consolidated Plastic Co.,
twinsburg, oH)
(Continued)
table 19.1 (Continued) packaging Materials for Foods processed by pulsed Electric Fields, high
pressure and Irradiation
processing packaging Method and
Method Food packaging Material Storage temperature Sources
hpp tomato 50 mL presterilized PP tubes Packed in a glove box, Min et al.
Juice, (Corning, Acton, MA) sanitized by H2o2 and uV (2003a,
orange storage at 4°C 2003b)
juice
Kimchi Pe bags Heat sealing without sohn and
(Korean entrapping any air bubbles. Lee (1998)
fermented storage in ice water
vegetable
product)
orange Plastic bags with eVoH storage at 0°C and 10°C takahashi
juice Pe/nylon pouches Vacuum sealing, storage at et al. (1998)
sliced 500 mL Pet bottle 3°C and 9°C Carpi et al.
cooked nylon/Pe bag (0.75 mil nylon, storage at 4°C (1999)
ham 2.27 mil Pe) (Koch supplies, Vacuum sealing, storage at Goodner
orange inc., Kansas City, Mo) 4°C et al. (1999)
juice 250 mL Pe bottle Chilled storage Murano et al.
Pork Plastic whirl-pak sampling bags Heat sealing. storage at (1999)
sausages (nasco, Fort Atkinson, Wi) 5°C, 15°C, and 25°C Ancos et al.
Low-fat Pe bags storage at 4°C and (2000)
yogurt Glass, PP, teflon, and barex 210 21°C–23°C Palou et al.
Guacamole (modified acetonitrile-methyl Flasks were closed by (2000)
salsa acrylate copolymers) flasks or screw lids. Pe pouches raghubeer
orange Pe pouches were heat sealed. storage et al. (2000)
juice, Laminated Pe-PA foil at 4°C Garcia et al.
orange- (Multiseven-80) storage at 4°C (2001)
lemon PA/Pe/eVoH/Pe, PA/PP/eVoH/ storage at 4°C tuboly et al.
carrot juice PP, Pe/PA/Pe, Pet/Al/Pe films (2003)
turkey meat (soplaril, elf-Atochem, Dax, Cruz et al.
Fatty duck France) (2003)
liver
Irradiation Potato steel tray, covered with a plastic Controlled atmosphere Ziegler et al.
Precooked Film storage (1968)
lobster 3 mil mylar/saran/Pe bags, (0.03, 0.5, 5, and 15% (v/v) Dagbjartsson
Chicken wrapped in Pe bags Co2 in air) and solberg
meat Air packaging: LDPe. Vacuum storage at 3°C (1973)
Pork patties, Packaging: unplasticized bags storage at 5°C Calenberg
meats, of PA and Pe (uPA/Pe 15_60, storage at 4°C et al. (1999)
turkey sudpack Verpackungen, storage at 4°C Ahn et al.
breast Germany) storage at 4°C (1998)
meat Air packaging: Pe bag (2 mil, Kim et al.
Cooked Associated bag Company, (2002)
pork Milwaukee, Wi). Vacuum nam and
sausages Packaging: nylon/Pe (Koch, Ahn (2002)
Ground Kansas City, Mo) Jo et al.
beef nylon/Pe bag (Koch, Kansas (2000)
patties City, Mo) Lopez-
(1) nylon/Pe bags (Koch Gonzalez
supplies, inc., Kansas City, Mo), et al. (2000)
consisting of 0.75 mil nylon and
2.25 mil Pe, with a moisture
transmission rate of 0.73 g/100
in 24 h atm and oxygen
permeability of 3.9 cc/100 in2
24 h atm. (2) saran/polyester/
Pe bags (Koch supplies, inc.,
Kansas City, Mo), consisting of
a top layer of 0.48 mil saran1
Source: Adapted from Min, C. s. et al., Innovation in Food Packaging, pp. 516–531, 2014.
volume of foods is a function of pressure applied, while an equal expansion occurs after decom-
pression; so, the packaging material used for HPP must be able to accommodate 15% reduction in
volume and regain to its original shape without any change in seal integrity and barrier properties.
Plastic films are generally suitable for HPP processing; however, it is not suitable for high tempera-
ture. Glass material and metal cans are generally not suitable for the HPP. Since air and gases are
very compressible under high pressure, the more the head space, the more will be strain deforma-
tion in packaging material; thus, the head space must be kept as low as possible (Lambert et al.,
2000). Various flexible materials including polypropylene (PP), polyester tubes, nylon PP pouches,
and polyethylene pouches (PE) are currently used as packaging material for HPP treatment or HPP-
treated foods. HP treatment adversely affected the barrier property of aluminium (Al) layer in multi-
layered film, which could be related to the promotion in rupture of less elastic material (Caner et al.,
2004; Min and Zhang, 2007). Dobias et al. (2004) studied the effect of HP treatment on physical
and chemical properties of polymer films and reported that the effect of HP on functional property
of single film was comparatively more than to the laminates. The tensile property of PE/polyamide
films were remaining unaffected after pressure treatment however up to 8%–17% increase in tensile
strength (cross direction) was observed in films after treatment of HPP. The water vapour perme-
ability of the majority of the films was increased after HP treatment; however, laminates of PE/PA/
EVOH/PA showed a decrease in water vapour permeability after pressure treatment. Failure in the
integrity of packaging material after HPP is very crucial for the food safety, so several researchers
conducted the study to know the effect of HPP on the physical and barrier properties, delamination,
and sealing properties of packages (Table 19.2).
table 19.2 Effect of hpp processing on properties of packaging Material
packaging
property Material hpp Conditions Effect references
barrier property PP/eVoH/PP, 400–600 MPa no significant Masuda et al. (1992)
oPP/PVoH/Pe, 600–800 MPa, 5, change in Caner et al. (2000)
KoP/CPP, Pet/ 10, 20 min, 45°C permeability of Lambert et al.
Al/CPP 200, 350 and o2 and water (2000)
Pet/siox/ 500 MPa vapour Lambert et al.
polyurethane 400 MPa, 30 min, no significant (2000)
(Pu) adhesives/ 60°C change in Caner et al. (2000)
LDPe, Pet/ 600–800 MPa, 5, permeability of Gallotta et al. (2009)
Al2o3/Pu/LDPe, 10, 20 min, 45°C o2 and water
Pet/PVDC/ 500 MPa, 15 min, vapour
nylon/HDPe/Pe, 50°C 800 MPa, 2 min,
Pe/nylon/eVoH/ 25°C
Pe, Pe/nylon/Pe, no significant
Pet/eVA, PP/PA/ change in water
Pe, PA/Pe, Pet/ vapour
PVDC/Pe, PA/ significant
PP/Pe increase in o2,
Pe coated with Co2 and water
siox and PP vapour
LLDPe/eVA/ permeability
eVoH/eVA/ significant
LLDPe, Pet/Al/ increase in o2,
PP Co2 and water
Metallized Pet/ vapour
eVA/LLDPe permeability
PetAlox
PLAsiox-PLA
(Continued)
table 19.2 (Continued) Effect of hpp processing on properties of packaging Material
packaging
property Material hpp Conditions Effect references
Mechanical property Pet/sioX/LDPe, 600–800 MPa, 5, no significant Caner et al. (2004)
Pet/Al2o3/LDPe, 10, 20 min, changes in Caner et al. (2004)
Pet/PVDC/ 45 min mechanical Gallotta et al. (2009)
nylon/HDPe/Pe, 800 MPa, 10 min, properties
Pe/nylon/eVoH/ 60°C no significant
Pe, Pe/nylon/Pe, 500 MPa, 15 min, changes in
PP/nylon/PP, 50oC mechanical
Pet/eVA/Pet 500 MPa, 15 min, properties and
LLDPe/eVA, 50°C tensile properties
eVoH/eVA/ no significant
LLDPe, Pet/ change in
PVDC/Pe, PA/ modulus of
Pe, surlyn, PA/ elasticity, tensile
PP/Pe strength
PetAlox increased, %
PLAsiox-PLA elongation
decreased
significantly
Volatile migration PP, Pe/nylon/ 800 MPa, 10 min, no significant Caner et al. (2004)
eVoH/Pe 60oC absorption of Masuda et al. (1992)
LDPe, eVA 400 MPa, 10 min D-limonene Goetz and Weiser
LDPe/HDPe/ 50 MPa 23°C Decrease in the (2002)
LDPe 150, 300, sorption of shauwecker et al.
Pe/nylon/Al/PP 600 MPa, 30 s, D-limonene (2002)
pouches 30°C, 50°C, 75°C Decrease in the
permeation rate
of p-cymene to
LDPe/HDPe/
LDPe
Decrease in the
lamination of
1,2 propanediol
(PG)
Delamination PA/Pe, Pet/ 200, 350, no delamination Lambert et al.
PVDC/Pe, PA/Pe 500 MPa, 30 min Delamination (2000)
surlyn, PA/PP/Pe 500 MPa, 300 s Delamination Goetz and Weiser
PA/Pe 200, 690 MPa (2002)
Pe/nylon/Al/PP shauwecker et al.
(2002)
sealing strength PP/eVoH/PP, no significant Masuda et al. (1992)
oPP/PVoH/Pe, change in heat
KoP/CPP, Pet/ seal strength
A1/CPP
Source: Adapted from Min, s. and Zhang, Q. H., Packaging for Non Thermal Processing of Food, pp. 70–71, 2007.
Packaging material with 12% deviation of mechanical and barrier property after HPP treatment
was considered as an acceptable limit for the packaging materials (Table 19.2). Flexible material
made up of ethylene vinyl alcohol (EVOH) and polyvinyl alcohol (PVOH) were found to be compat-
ible with HPP treatment (Masuda et al., 1992). Water barrier and oxygen barrier property of several
laminated plastic films (like PP/EVOH/PP, oriented polypropylene (OPP)/PVOH/PE, KOP/cast
polypropylene (CPP), and polyethylene terephthalate (PET) remain unaffected when treated up
to 600 MPa (Ozen and Floros, 2001). Oxygen permeability of the PE coated with SiOx and PP
materials was also not changed by a treatment of HP at 800 MPa (2 min, 25°C) (Lambert et al.,
2000); however, a significant increase in the permeability of O2, CO2, and water vapor was observed
in a metalized PET film after an HP treatment (Caner et al., 2000, 2004).
Mechanical properties of the film were not negatively affected by HPP treatment (Caner et al.,
2004); however, tensile strength of PE, BOPP, PET/polyvinylidiene chloride, PA/PE surlyn, and
PA/PP/PE were increased after HP treatment, indicating that the material became rigid and less
flexible after pressure treatment but all the changes were below the 25% (Lambert et al., 2000;
Dobias et al., 2004). Galotta et al. (2009) also observed the large reduction in % elongation in
PETAlOx film when exposed for 15 min at 50°C under 500 MPa. Significant reduction in % elonga-
tion was due to the formation of pinholes and cracks in the structure during HPP, acting as stress
concentrators and causing early failure of film (Gallotta et al., 2009). They further explained that
during HPP treatment increased heat of fusion induced crystallization and the crystallites reinforce
the polymeric structure and so increased the tensile strength of PETAlOx and PETAlOx-PLA film.
In PETAlOx-PLA film, no significant change was observed in modulus of elasticity due to exposure
of HPP. As per Le-Bail et al. (2006), modulus demonstrates the firmness of the structure and gener-
ally the higher the degree of crystallinity, the higher is the modulus.
During compression, free volume of packaging material decreases and it decreases the capac-
ity for absorption of flavouring compounds from food (Caner et al., 2004), and when the pressure
is released packaging material quickly recovers its original volume and thus sorption and diffu-
sion proceeds at the same rate as expected (Caner et al., 2004). Some packaging material failed to
recover their original volume, reducing the migration of flavouring compound from the food to the
material. Several authors reported that the sorption of aroma compound was comparatively lower
in pressurized films than in non-pressurized films (Masuda et al., 1992; Kubel et al., 1996). Kubel
et al. (1996) suggested that the transition of the film to the glassy state under high pressure may be
one of the reasons for the decrease in sorption of flavouring compounds in the material.
Sealing is one of the major critical parameters for producing the HPP-treated foods, and it should
be critically investigated because leakage may occur through sealed areas after HPP (Caner et al.,
2004). There is very little in the literature on the effect of HPP on sealing integrity of the packaging
material. Sealing integrity of the copolymers (PE/PA/EVOH/PE and PET/PE/PVOH/PE) was not
significantly changed after HPP treatment (200–500 MPa, 15 min) (Masuda et al., 1992); however,
sealability of copolymer of ethylene and methacrylic acid was increased after treating with HPP at
600 MPa for 60 min (Dobias et al., 2004).
19.3 IrraDIatION
The irradiation of food is considered as a food preservation technique and federal and Food
Drug and Cosmetic Act (the Act) defines the use of irradiation to treat or inspect the food as a food
additive which is subject to regulation by the U.S. Food and Drug Administration (FDA). The Act
defines a food additive as: “any substance the intended use of which results or may reasonably
be expected to result, directly or indirectly, in its becoming a component or otherwise affecting
the characteristics of any food…including any source of radiation intended for such use…”. All
irradiation processes must obtain approval from the FDA as FDA irradiations are defined as food
additives. This includes packaging material also, because packaging component could become part
of food by migration from packaging to food (Komolprasert, 2016). The USFDA has approved
number of packaging material for use in packaged food for irradiation (USFDA, 1989). The Health
and Welfare Agency of Canada granted FDA approval for the irradiation of poultry at the level of
1.5–3 kGy in 1990; similarly, in 1997 FDA approved the use of irradiation in beef, lamb, and pork.
A dose of 1.0 kGy is recommended in ground beef to minimize the losses of sensory attribute
(Morris et al., 2006). More than 40 countries have approved irradiation in more than 100 foods. The
Joint FAO/IAEA/WHO Expert Committee ensured that application of irradiation up to 10 kGy does
not produce any toxicological or microbiological hazards in foods (Lee et al., 2004). The group con-
cluded that high-dose irradiation, conducted in accordance with good manufacturing and irradiation
Figure 19.1 image of internationally recognized logo of irradiated food.
practices, could be applied to several types of foods to improve their hygienic quality, to make them
shelf-stable, and produce special products (WHO, 1999). Labelling for irradiated food is required
and an international logo (Figure 19.1) must also appear on the package and the following statement,
“treated by irradiation,” should be displayed (Lacroix et al., 2002).
The use of irradiation for decontamination of a food is an impressive technology which can
be applied to end food product. This technology is also applicable to fresh, frozen, and cooked
products. Irradiated foods are the foods which have been treated with the ionizing radiation where
foods are exposed with the direct action of electronic, electromagnetic rays to ensure the innocuity
of food and to enhance the shelf life of foods (Lacorix, 2014). When X rays and gamma rays
bombard the food materials, they can knock off an electron from atoms and molecules within it,
causing ionization so that it is called ionization radiation (Lacorix et al., 2002). Co-60 (1.17 and
1.33 MeV) and Cs-137 (0.662 MeV) are the permitted radiation sources in food. Gamma rays
and X-rays transfer the energy in so many ways; each involves the liberation of fast electrons that
then lose energy by electronic interactions. Another source of irradiation is β-rays which are the
beam of electrons having maximum 10 MeV energy. The treatment received by the food product is
characterized by the irradiation dose, which is the quantity of energy absorbed by the food. The unit
of absorption of dose is the gray (Gy). Depending upon the dose applied the food may be pasteur-
ized to inactivate pathogenic microorganism or they may be sterilized to eliminate all microorgan-
isms (Olson, 1998). Irradiation inactivates microorganisms by disrupting their nucleic acid (DNA)
through the formation of pyrimidine dimers between adjacent pyrimidine molecules on the same
strand of DNA (Grecz et al., 1983; Franz et al., 2009) and so preventing the microorganisms from
replicating. Irradiation can enhance shelf life by reducing putrefactive bacteria and by inactivating
specific pathogens like Bacillus cereus, Cronobacter sakazakii, S. aureus, E. coli, and Salmonella
typhimurium (Franz et al., 2009). Radiations also damage the other components of microorganisms
like membranes, cells, and plasmids (Smith and Pillai, 2004). The size of the DNA is one of the
most important factors in irradiation. Parasites and insects, which have a large amount of DNA, are
rapidly killed by very low doses of irradiation of 0.1 kGy or less. Bacteria are the more irradiation
tolerant due to smaller DNA with D values in the range of 0.3 to 0.7 kGy. As per Berry (2004),
irradiation is the most effective technique to eliminate pathogenic microorganisms from the food
supply. Spore-forming bacteria are the generally most heat resistant with D values in the order of
2.8 kGy (Lacorix, 2014), which is due to the low moisture content (less than 10% on wet basis) in
comparison to vegetative bacteria (Dickson, 2001). The prion particles which are associated with
bovine spongiform encephalopathy do not have nucleic acid, so they are not generally inactivated
by irradiation (Young, 2003).
The amount of vitamin loss due to exposure of radiation depends on the dose of irradiation,
temperature, presence of oxygen, and type of foods. In general, irradiation at low temperature and
in absence of oxygen minimizes the loss of vitamins during treatment (Olson, 1998).
19.3.1 packaging for Irradiation
Irradiation is commonly used for sterilization of packaging material for aseptic packaging in
foods and pharma (Ozen and Floros, 2001). Foods are generally pre-packaged before irradiation to
avoid any cross contamination. Packaging materials authorized for some countries for food irradia-
tion and for packaged foods are given in Tables 19.3 and 19.4, respectively. The effect of irradiation
on packaging material is well established; however, the effect of irradiation on packaging material
like laminates, coextruded, etc. still needs further investigation. The two main effects of irradiation
on flexible packaging are cross-linking and chain degradation; these two effects occurs simultane-
ously and the polymer composition, condition of irradiation, and dose decide which effects will
dominate. Vacuum or inert atmosphere during irradiation favours the cross linking of polymers;
however, chain scission governs during irradiation in the presence of oxygen and air (Chuaqui-
Offermanns, 1989; Ozen and Floros, 2001; Komolprasert and Morehouse, 2004; Han, 2007). The
irradiation of polymeric materials leads to the development of very reactive ingredients, i.e., free
radicals and ions, which are very unstable in nature and can propagate several undesirable reactions
resulting in disproportion, hydrogen abstractions, arrangements, and formation of new bonds. The
degree of these deformations varies with structure of the polymer and the conditions of polymer
before and after treatments (Chmielewski et al., 2005). Killoran (1983) investigated the safety of
tin plate cans after subjection to irradiation dose. Packages were sound and safe after exposure of
75 kGy. Normal glass may discolour after exposure to irradiation, so glass may not be good choice
for irradiation. Glass packaging material used for medical application is specially formulated so it
does not tint when exposed to irradiation (Food Safety, 2005). Killoran (1983) studied the effect of
irradiation of 71 kGy on five different types of polymer pouches and he suggested that the irradia-
tion can easily be adapted for use with flexible films. He also observed that quality of irradiated
meat remained unaffected and was found to be acceptable for two years when stored at 21°C tem-
perature. PE, polystyrene, and PET were observed to be well suited for practice with food intended
for irradiated. Satisfactory irradiation-resistant packaging material from polymers can be prepared
by using co-extrusion techniques (Thayer, 1988).
Killoran (1983) studied the effect of exposure of gamma radiation of 100 kGy on the optical
properties of PE “LDPE” amide 6-amide 6.6 copolymer “PA6-PA6.6” and PET. A marked reduc-
tion in % transmittance (at low wavelength) was reported in all the tested films except PET where
no significant changes were observed in either transmittances or light absorption (Moura et al.,
2004). Table 19.5 presents the effect of irradiation on barrier properties of films. As per Han et al.
(2004), exposure of irradiation dose of 1.5 and 3.2 kGy improved the oxygen barrier capability of
LDPE films by 7.7% and 4.5%, respectively; however, irradiation did not show any obvious effect on
water barrier property and stiffness of LDPE films. Rojas De Gante and Pascat (1990) reported that
the 25 kGy of irradiation dose did not change oxygen permeability of LDPE and OPP films. Pilette
(2003) also did not find any changes in the oxygen and water vapour permeability of PE.
Irradiation did not affect water vapour permeability and stiffness of LDPE film (Han et al.,
2004). Radiation doses of up to 30 kGy had no significant effect on water vapour permeability and
table 19.3 packaging Materials Specifically authorized for Food Irradiation in Some
Countries
packaging Material Country Where Specifically authorized

Cardboard Poland, united Kingdom
ethylene-vinyl acetate copolymer usA
ethylene-vinyl acetate coextruded Canada, usA
Fibre board, wax-coated Canada, usA
Fibre board india
Glass india
Glassine paper usA
Hessian united Kingdom
Kraft paper usA
nitrocellulose coated cellophane india, usA
nylon 6 india, usA
nylon 11 india, usA
Paper Poland, united Kingdom
Paperboard, wax-coated india
Paper, coated (hot-melt) Poland
Paper, coated (polyethylene) Poland
Paper/aluminium foil laminates Poland
Paper/aluminium foil/ionomer laminates Poland
Polyamide Poland
Polyamide-polyethylene Poland
Polyester-metallized-polyethylene Poland
Polyester-polyethylene Poland
Polyethylene india, usA
Poly(ethylene terephthalate) india, usA
Polyethylene (extensible) Poland
Polyethylene (high-density) Poland
Polyethylene (low-density) Poland
Polyethylene/paper/aluminium foil laminates Poland
Polyolefine usA
Polyolefine (high-density) Canada
Polyolefine (low-density) Canada
Polypropylene Poland, united Kingdom
Polypropylene metallized Poland
Polystyrene Canada (as foam), india, usA
rubber hydrochloride india, usA
steel, tin-plated or enamel-lined india
Vegetable parchment india, usA
Vinyl chloride-vinyl acetate copolymer india, usA
Vinylidine chloride copolymer-coated cellophane usA
Vinylidine chloride-vinyl chloride copolymer india, usA
Wood india, Poland
“Viscosa” Poland
Source: Adapted from Chmielewski, A. G., Packaging for food irradiation. rAPorty iChtJ.
seriA b nr 1/2006, 2006, http://www.iaea.org/inis/collection/nCLCollection store/_
Public/38/005/38005202.pdf.
table 19.4 packaging Materials approved for the Use in Irradiation of pre-packaged Foods
Canada
Commercial Designation Composition Date of acceptance
eD 100/8 Polyolefin (high-density) outer layer polyolefin June 30, 1989
(high-density) middle layer polyolefin
(high-density) sealant layer’
Polystyrene foam trays styron 685D June 30, 1989
Film ssD 300; film ssD 500, D 900 Polyethylene and ethyl vinyl acetate coextruded May 20, 1988
bags: L 300, L 500, L 600, Polyethylene and ethyl vinyl acetate coextruded July 20, 1988
L300
boxes (contact, no contact) Fibre board wax-coated June 30, 1989
Source: radiation Applications research branch, AeCL, Canada, adapted from Chmielewski, A. G., Packaging
for food irradiation. rAPorty iChtJ. seriA b nr 1/2006, 2006, http://www.iaea.org/inis/collection/
nCLCollectionstore/_Public/38/005/38005202.pdf.
table 19.5 Effect of Irradiation on properties of packaging Material

packaging
property Material Irradiation Dose Effect references
barrier properties LDPe, HDPe, 8 kGy no significant changes in Crook and boylston
Pet, PVC 25 kGy gas permeability (2004)
Pe pouch 5, 10, 30 kGy no significant changes in rojas De Gante and
eVA, HDPe, Ps, (γ-radiation) the permeability of o2 Pascat (1990)
boPP, LDPe and vapour Goulas et al. (2002)
no significant changes in
Co2, o2 and water
vapour
Mechanical Ps 100 kGy no changes Pentimalli et al.
properties PP, LDPe, PLA (γ-irradiation) no changes (2000)
eVA, HDPe, Ps, 0.5–2 kGy no significant change in Krishnamurthy et al.
boPP, LDPe 5, 10 kGy tensile strength, % (2004)
HDPe, boPP 30 kGy elongation at break, Goulas et al. (2002)
LDPe 30 kGy young’s modulus Goulas et al. (2002)
tensile strength Goulas et al. (2002)
decreased
% elongation at break
decreased
Volatile formation LDPe, oPP <25 kGy H2o2 carbonyl rojas De Gante and
PP 40 kGy compounds (Ketones Pascat (1990)
Pet 25 kGy and aldehyde) Marque et al. (1995)
Alkyl radicals Komolprasert et al.
Formic acid, acetic acid, (2001)
1,3 dioxalane, 2 methyl
1,3 dioxalane
sealing strength eVA Decrease in heat seal Matsui et al. (1991)
surlyn strength Hog and
no significant change in Cumberbatch
heat seal strength (1991)
Source: Min, s. and Zhang, Q. H., Packaging for Non Thermal Processing of Food, pp. 70–71, 2007.
gas permeability (O2 and CO2) of EVA, HDPE, BOPP, LDPE, or PS (Goulas et al., 2002). With
increasing radiation dose, the diffusion coefficient of EVA film is increased due to chain scission in
EVA film (Matsui et al., 1991).
Effect of exposure of 50 kGy irradiation on mechanical properties of two types of Surlyn film
was investigated by Hog and Cumberbatch (1991). Mechanical properties including tensile strength,
% elongation, modulus, and tear strength of exposed film were not changed significantly and heat
seal strength was also maintained over a period of one year. However, heat seal strength of irradiated
EVA film decreased with increase in dose of irradiation (Matsui et al., 1991). Mechanical strength
of PS film was not changed significantly when exposed to the 100 kGy irradiation (Table 19.5). The
effect of gamma irradiation doses of up to 5–30 kGy on monolayer flexible packaging film, EVA,
HDPE, PS, BOPP, and LDPE was studied by Goulas et al. (2002). Tensile strength, percentage elon-
gation at break, and Young’s modulus of all the films were not changed significantly after absorbed
doses of 5 and 10 kGy. The tensile strength of BOPP and HDPE films decreased after exposure of
30 kGy (Table 19.5).
Irradiation (mainly the higher dose of 60 kGy) induced certain small, but statistically signifi-
cant, differences in the mechanical properties of multilayer films (with or without recycled LDPE
layer), while no significant differences were found in the thermal properties, gas, and water vapour
permeability of multilayer films. The above findings are discussed in relation to the good quality of
the pre-consumer scrap used in the present study (Chyitri et al., 2005).
Generally, flexible packaging material produces a number of volatile components when exposed to
irradiation. EI Makhzoumi (1994) reported that 63 different volatiles were produced from PE, PET, and
OPP (El) after irradiation. Low molecular weight volatiles products of LDPE and PP were investigated
by thermal de-sorption spectroscopy gas chromatography-mass spectrometry after exposure of 25 kGy.
Hydrocarbon, aldehydes, ketones, and carboxylic acids were produced after exposure of irradiation
(Buchalla et al., 1993). Komolprasert et al. (2001) reported that formic acid, acetic acid, 1,3-dioxalane,
and 2-methyl-1,3-dioxalane were the major volatiles which were produced from the PET films after
exposure of 25 kGy. A 25 kGy irradiation dose was sufficient to produce volatiles component from
crystalline and oriented semi-rigid PET film (Komolprasert et al., 2001). LDPE, HDPE, PVC, and PET
can release hydrogen, carbon dioxide, carbon monoxide, and methane gases on irradiation and can also
release volatile oxidation products including peroxides, alcohols, aldehydes, ketones, and carboxylic
acids (Crook and Boylston, 2004). The types and concentration of volatiles produced were increased
with increase in the dose of irradiation. The produced volatiles and non-volatiles during irradiation
affected the sensory properties of potable water after coming in contact with packaging films. Change
in taste of water was observed mainly at higher side (Chyitri et al., 2005).
19.4 pULSED ELECtrIC FIELD
PEFs have been developed to achieve sufficient microbial inactivation where the use of electric
field for few seconds causes local structural changes and rapid breakdown of cell membrane (Toepfl
et al., 2005). PEF is a non-thermal technique which uses the short pulses of electricity to inactivate
the microorganism without any detrimental adverse effect on food quality attributes. For food qual-
ity attributes, PFE technology is considered superior to traditional thermal technology as it reduces
the detrimental changes in chemical, physical, and organoleptic attributes of food (Quass, 1997).
During PEF treatment a pore is developed in cell membrane, which can be reversible or irrevers-
ible. In the field of food technology, irreversible pore formation of PFE is used as mild preservation
technique for liquid food (such as milk, yoghurt, juices, liquid egg, brines, soups, etc.) and also as
a substitute of conventional techniques for cell disintegration (Toepfl et al., 2005). PEF inhibits
microorganisms by breaking the microbial cells (Devlieghere et al., 2004), which leads to leakage
of ions, losses of metabolites, protein release, and increased uptakes of drugs and molecular probes
(Kinosita and Tsong, 1977; Benz and Zimmermann, 1980). PEF technology had been extensively
used by several scientists and numerous studies demonstrated the efficiency of PEF to produce
shelf-stable plant-based liquid foods with better retention of nutritional and organoleptic qualities
(Odriozola-Serrano et al., 2013; Saldaña et al., 2014). Moreover, high electric field pulses can also
be used as pre-treatment of solid food matrices to improve processes such as extraction of peptides
and polysaccharides, osmotic dehydration, drying, and freezing (Donsi et al., 2010). PEF has very
limited effect against microbial spores and cannot be used for products containing air or which can
form air bubbles, and also not applicable for foods having high or variable electrical conductivity.
Critical factors that can affect the inactivation of microorganisms during PEF treatments are
process variables, media, and microbial factors. Process variables include pulse wavelength and
width, electrical field intensity, temperature, and time of pulse exposure (Hulsheger and Niemann,
1980; Zhang et al., 1994, 1995; Barbosa-Canovas et al., 1998; Clark, 2006). The most important
critical parameters of a product include electric conductivity, pH, density, viscosity, and water
activity (Zhang et al., 1995; Vega-Mercado et al., 1996; Wouters et al., 2001; Min et al., 2003). The
type of microorganism, cell, size and shape of microorganism, growth stage, and microbial stress
adaptation also influence the efficiency of PEF inactivation (Grahl and Markl, 1996; Raso et al.,
1998; Wouters et al., 2001; Evrendilek and Zhang, 2003). Researchers showed that increasing the
pulse number (n) increased the number of cellular injuries (Hodgins and Mittal, 2003).
19.4.1 packaging requirement for pulse Electric Field
Aseptic packaging is considered the most appropriate way for packaging of PEF-processed food
products (Qin et al., 1995; Ayhan et al., 2001). Shelf life of PEF-processed products stored in plastic
packaging material was studied by several scientists. Table 19.1 presents the list of packaging materi-
als for foods processed by PEFs. The shelf life of food stored in packaging material depends on the
permeation of water and gas through packing material because a significant amount of spoilage of food
results from oxidation and changes in the moisture content of food (Min and Zhang, 2007). Plastic- and
paper-laminated materials are widely used as packaging material for aseptic packaging. The materials
are usually thermo-formed to produce food containers for aseptic processing. The thermo-forming
conditions should be optimized before aseptic PEF processing so that the material used does not lose
its barrier and mechanical properties. Ayhan et al. (2001) studied the effect of packaging material,
storage temperature, and storage time on the quality attribute of PEF-processed orange juice. The
orange juice was treated with PEF at an electric strength of 35 KV/cm for 59 µs. Flavour, colour, and
vitamin C retention of PEF-treated orange juice in four different packaging materials, i.e., sensitized
glass, PET, LDPE, and HDPE were investigated. Glass and PET bottles were comparatively effective
at retarding degradation of flavour, colour, and vitamin C of PEF-treated orange juice during storage at
4°C for 112 days than to orange juice stored in HDPE and LDPE, which might be due to relatively low
barrier property of polythene against oxygen. The degradation of lycopene in PEF-processed tomato
juice stored in PP tubes were more significant during the first seven days of storage at 4°C (Min et al.,
2003). Carotenoids are degraded in foods through oxidation (Thakur et al., 1996) and, thus, the signifi-
cant reduction was considered due to availability of oxygen in the head space of PP tube (Min et al.,
2003). Modified atmospheric packaging (MAP), which limits the availability of oxygen in head space,
may be applied to reduce the rate of oxidation in PEF-processed food (Min and Zhang, 2007).
19.5 pULSED LIGht
PL is an emerging non-thermal technique for decontamination, which is capable of inactivat-

ing microorganisms on various surfaces including food, equipment, and food packaging material
(Dunn et al., 1995). PL technology comprises the generation of high-power electrical pulses that
are subsequently changed into short-duration, high-power pulses of broad-spectrum (180–1100 nm)
electromagnetic waves via an inert-gas (mainly xenon) flash lamp. As per Dunn et al. (1989) the
shorter the pulse duration, the higher the delivered energy and consequently the inactivation of
microorganism. Heinrich et al. (2015) provided the schematic layout of a PL device. PL lamp pro-
duced light of a broad spectrum ranging from UV radiation to near-infrared radiation. The light
used for food application is usually pulsed at 1–20 flashes per second and the energy density at the
surface is in the range of 0.01–50 J/cm2 (Condon et al., 2014). PL sterilization technique is an easy
operation and short irradiation duration. Traditional chemical sanitizers (H2O, peracetic acid, etc.)
used for sterilizing the surfaces can be replaced by PL technology. The FDA has approved the use
of PL technology in the production of food, processing, and handling (Code of Federal Regulation,
CFR: 21CFR179.41). Microbes exhibit variation in PL susceptibility and show the following trends
in decreasing order: gram-negative bacteria, gram-positive bacteria, fungal spores, and bacterial
spores (Oms-Oliu et al., 2010; Levy et al., 2012). The colour of fungal spores can play an important
role in susceptibility and Aspergillus niger spores are more resistant than Fusarium culmorum
spores (Esbelin et al., 2013). The antibacterial property of UV light is basically due to its absorp-
tion by conjugated double bonds in protein and nucleic acid of microorganisms. PL was found to be
significantly more effective against microorganisms than continuous ultraviolet lightening system
(Barbosa-Canovas et al., 1998).
The food matrix affects the efficiency of PL in regards to transparency, opacity, surface charac-
teristics, and composition. Low reflection, high absorption and transmission coefficients of matrix
are helpful in achieving optimal results (Dunn et al., 1989; Palmieri and Cacace, 2005; Gómez
et al., 2007; Robert et al., 2017).
The surface of food matrix should be as smooth as possible; immense irregularities and light
absorbing material constitutes an accommodation for microbial contamination and a hindrance for
incident light (Dunn et al., 1995; Gómez-López et al., 2005a, 2007; Palmieri and Cacace, 2005;
Lagunas-Solar and Gómez-López, 2006; Sommers et al., 2009). Food matrix should contain very
low quantity of substances able to absorb light such as protein and fat; however, carbohydrates
do not show this noticeable light absorbing effect (Gómez-López et al., 2005a; Rajkovic et al.,
2010). Physiological distribution, population, density distribution, and growth parameter (growth
rate and lag time) also affect the efficiency of PL (Dunn et al., 1989; Augustin et al., 2011; Cudemos
et al., 2013).
19.5.1 packaging requirement for pulsed Light treatment
Packaging material used for PL treatment should be transparent for light transmission (Han,
2007). Packaging material used for irradiation should be resistant to heat because PL treatment can
result in excessive heat generation at the surface of the food (Kumar and Han, 2012). Those packag-
ing materials which cannot tolerate autoclave temperature are not recommended for PL sterilization.
The resistant to heat of polyvinyl chloride (PVC) makes it a most preferable packaging material for
PL treatment (Katz, 1999). For satisfactory in-package results of PL appropriate selection of pack-
aging materials with regards to transmissibility of light is required. Opaque packaging material and
matrices that may interfere with absorption of light should be avoided (Dunn et al., 1989; Palmieri
and Cacace, 2005; Elmnasser et al., 2007; Han, 2007; Oms-Oliu et al., 2010; Heinrich et al., 2015).
Since metals (tin, aluminium, tin-free steel, etc.), paper, and paperboard offer excellent pro-
tection against light, glass, and polymeric material (plastics) can only be considered as preferred
packaging material in package application of PL (Dunn et al., 1989; Eie, 2009). Depending on the
type of glass, its transmittance for the wavelength of UV range is limited and may therefore limit
the application of glass in PL effect (Eie, 2009). Out of more than 30 types of polymers available, the
most common polymers used in food packaging are polyolefins, PP, polyesters, polycarbonate (PC),
or polyethylene naphthalate (PEN) (Heinrich et al., 2015). Bio-based polymers (e.g., polylactic acid
(PLA)) are also becoming increasingly important in the food industry (NOVA Institute, 2013).
Depending on the basic structure and processing characteristics a polymer material may have a
wide range of physical, chemical, and mechanical characteristics (Kirwan and Strawbridge, 2003).
Transmittance of polymeric material is strongly influenced by some of the additives that are added
for market appeal or any functional property during processing (Crompton, 2007; Pospíšil and
Nešpůrek, 2008). Ultraviolet protective agents used as additives can be UV screens or substances
which may interfere with PL light.
Till now, only a few authors studied the effect of PL on food-contact materials. While the
emphasis was laid on the physical stability and mechanical stability, little concern was paid on
chemical movement and safety concerns. But awareness about the effects of PL exerted on the
packaging materials is on the rise (Ozen and Floros, 2001; Han, 2007; Guillard et al., 2010; Castillo
et al., 2013). Keklik et al. (2009, 2010) studied the effect of PL treatment on mechanical proper-
ties of the PP material in both along-machine and perpendicular-to-machine directions. Elastic
modulus, yield strength, and percent elongation were recorded before and after the treatment.
In both studies, authors suggested that PL may have more impact on the elastic modulus and yield
strength compared to tensile strength and percent elongation at yield or break point. The described
changes were more prominent in moderate and extreme PL treatment conditions (Keklik et al.,
2009, 2010). Ringus and Moraru (2013) investigated the effect of PL treatment on the structural
and physical properties of surface-treated low- and high-density polyethylene (LDPE and HDPE),
namely MET, TR and EP. Surface hydrophobicity through water contact angle, to indicate changes
in surface structure, water barrier property, and bacterial adherence, had also been measured by
several authors (Raab et al., 1982; Mafu et al., 1991; Bower et al., 1996; Li and Logan, 2005; Dury-
Brun et al., 2007). Surface roughness was tested to determine the possibility of microorganisms
shading or hiding from the PL treatment. The findings showed that the most reflective materials,
MET, EP and TR, also showed the highest roughness and, hence, were last in allowing surface
sanitization (Woodling and Moraru, 2005; Ringus and Moraru, 2013). PL only had a minimal
impact on water-contact angles and, therefore, has no significant adverse effect on structural and
physical properties of PL-treated film (Ringus and Moraru, 2013). Although Keklik et al. (2009,
2010) and Ringus and Moraru (2013) did not observe drastic alterations in the polymeric packaging
materials, it is always recommended to perform PL treatment within reasonable limits (Heinrich
et al., 2015). The capability of PL for the fast generation of heat can be revealed on the basis of
the photothermal effect. The temporary overheating and then rupture of cells without increase in
temperature of the surrounding surface is unique to PL above specific treatment intensity and is
known as pulsed UV disintegration (Wekhof, 2000; Wekhof and Trompeter, 2001). The microor-
ganisms, unable to disperse the energy taken up during the PL treatment, face breakdown and are
even capable of melting into the surrounding matrix, provided a certain specific surface tempera-
ture is reached (Wekhof and Trompeter, 2001; Takeshita et al., 2003; Cheigh et al., 2012). Wekhof
and Trompeter (2001) observed the same phenomenon for Aspergillus niger spores via electron
microscopy.
19.6 CONCLUSION
Non-thermal processing technologies are fulfilling the consumer’s demands for fresh-like
food and also to provide microbial safety. Non-thermal processes offer shelf life extension of food
without the application of any preservatives or additives. Both effects of non-thermal processing
on packaging material and effect of packaging material on non-thermally processed food dur-
ing storage period should be investigated thoroughly for the proper selection of packaging mate-
rial. It is assumed that active packaging, MAP antioxidant, and edible coating would extend the
shelf life of foods during storage. Thus, it is suggested to study the combination of smart/active
packing and non-thermal processing in future. Future research should emphasize HPP/PEF/PL/
irradiation-induced alteration in food and packaging material for successfully commercialization of
non-thermal technology, particularly for packaging applications.
rEFErENCES
Ahn, D.U., Olson, D.G., Lee, J.I., Jo, C., Wu, C., and Chen, X. (1998). Packaging and irradiation effects on
lipid oxidation and volatiles in pork patties. J Food Sci. 63(1): 15–19.
Ancos, B., Cano, M.P., and Gomez, R. (2000). Characteristics of stirred low-fat yoghurt as affected by high
pressure. Int Dairy J. 10: 105–111.
Augustin, J.C., Bergis, H., Midelet-Bourdin, G., Cornu, M., Couvert, O., Denis, C., et al. (2011). Design of
challenge testing experiments to assess the variability of Listeria monocytogenes growth in foods. Food
Microbiol. 28: 746–754.
Ayhan, Z., Yeom, H.W., Zhang, H., and Min, D.B. (2001). Flavor, color, vitamin C retention of pulsed electric
field processed orange juice in different packaging materials. J Agri Food Chem. 49: 669–674.
Balasubramanian, S., and Balasubramaniam, V.M. (2003). Compression heating influence of pressure trans-
mitting fluids on bacteria inactivation during high pressure processing. Food Res Internat. 36(7):
661–668.
Barbosa-Canovas, G.V., Pothakamury, U.R., Palou, E., and Swanson, B.G. (1998). Non-Thermal Preservation
of Foods. New York: Marcel Dekker, pp. 9–52, 139–213.
Benz, R., and Zimmermann, U. (1980). Pulse-length dependence of the electrical breakdown in lipid bilayer
membranes. Biochimica Biophysica Acta. 597: 637–642.
Berry, D. (2004). Keeping foods fresh. In: Food Product Design. http://www.foodproductdesign.com/
archive/2004/0104De.html.
Bower, C.K., McGuire, J., and Daeschel, M.A. (1996). The adhesion and detachment of bacteria and spores on
food-contact surfaces. Trends Food Sci Technol. 7(5): 152–157.
Buchalla, R., Schuttler, C., and Bogl, K.W. (1993). Effects of ionizing radiation on plastic food packaging
materials: A review. Part 1. Chemical and physical changes. J Food Protect. 56: 991–997.
Calenberg, S.V., Philips, B., Mondelaers, W., Cleemput, O.V., and Huyghebaert, A.J. (1999). Effect of
irradiation, packaging, and post irradiation cooking on the thiamin content of chicken meat. J Food
Protect. 62(11): 1303–1307.
Caner, C., Hernandez, R.J., and Harte, B.R. (2004). High-pressure processing effects on the mechanical,
barrier and mass transfer properties of food packaging flexible structures: A critical review. Packaging
Technol Sci. 17: 23–29.
Caner, C., Hernandez, R.J., and Pascall, M.A. (2000). Effect of high-pressure processing on the permeance of
selected high-barrier laminated films. Packaging Technol Sci. 13: 183–195.
Carpi, G., Squarcina, N., Gola, S., Rovere, P., Pedrielli, P., and Bergamaschi, M. (1999). Application of
high pressure treatment to extend the refrigerated shelf-life of sliced cooked ham. Ind Conserv. 74(4):
327–339.
Castillo, R., Biedermann, M., Riquet, A.M., and Grob, K. (2013). Comprehensive on-line HPLC-GC for
screening potential migrants from polypropylene into food: The effect of pulsed light decontamination
as an example. Polymer Degradation Stab. 98: 1679–1687.
Cheftel, J.C. (1995). Review: High-pressure, microbial inactivation and food preservation. Food Sci Technol
Int. 1: 75–90.
Cheigh, C.I., Park, M.H., Chung, M.S., Shin, J.K., and Park, Y.S. (2012). Comparison of intense pulsed light
and ultraviolet (UVC)-induced cell damage in Listeria monocytogenes and Escherichia coli O157:H7.
Food Cont. 25: 654–659.
Chen, B.Y., Lung, H.M., Yang, B.B., and Wang, C.Y. (2015). Pulsed light sterilization of packaging materials.
Food Package Shelf Life. 5: 1–9.
Chmielewski, A.G. (2006). Packaging for food irradiation. RAPORTY IChTJ. SERIA B nr 1/2006.
http://www.iaea.org/inis/collection/NCLCollectionStore/_Public/38/005/38005202.pdf (Accessed on
March 14, 2017).
Chmielewski, A.G., Haji-Saeid, M., and Shamshad A. (2005). Progress in radiation processing of polymers.
Nuclear Instrum Methods Physics Res Section B. 226: 44–54.
Chuaqui-Offermanns, N. (1989). Food packaging materials and radiation processing of food: A brief review.
Internat J Radiation Applicat Instrument Part C. Radiat Physics Chem. 34(6): 1005–1007.
Chyitri, S., Goulas, A.E., Badeka, A., Riganakos, K.A., and Kontominas, M.G. (2005). Volatile and non-volatile
radiolysis products in irradiated multilayer coextruded food-packaging films containing a buried layer
of recycled low-density polyethylene. Food Additives Contam. 22(12): 1264–1273.
Clark, J.P. (2006). High pressure processing research continues. Food Technol. 60(2): 63–65.
Condon, S., Alvarez, I., and Gayan, E. (2014). Non-thermal processing-pulsed UV light. In: Encyclopedia of
Food Microbiology, pp. 974–981. Edited by C.A. Batt. New York: Elsevier.
Crompton, T.R. (2007). Additive Migration from Plastics into Foods: A Guide for Analytical Chemists.
Shawbury, England: Smithers Rapra Technology Limited.
Crook, L.R., and Boylston, T.D. (2004). Flavour characteristics of irradiated apple cider during storage: Effect
of packaging materials and sorbate addition. J Food Sci. 69: C557–C563.
Cruz, C., Moueffac, A.E., Antoine, M., et al. (2003). Preservation of fatty duck liver by high pressure treatment.
Int J Food Sci Technol. 38: 267–272.
Cudemos, E., Izquier, A., Medina-Martínez, M.S., and Gómez-López, V.M. (2013). Effects of shading and
growth phase on the microbial inactivation by pulsed light. Czech J Food Sci. 31(2): 189–193.
Dagbjartsson, B., and Solberg, M. (1973). Textural changes in precooked lobster (Homarus americanus) meat
resulting from radurization followed by refrigerated storage. J Food Sci. 38(1): 165–167.
Devlieghere, F., Vermeiren, L., and Debevere, J. (2004). New preservation technologies: Possibilities and
limitations. Internat Dairy J. 14: 273–285.
Dickson, J.S. (2001). Radiation inactivation of microorganisms. In: Food Irradiation, Principles and
Applications, pp. 23–35. Edited by R. Molins. New York: Wiley-Inter Science, John Wiley & Sons.
Dobias, J., Voldrich, M., Marek, M., and Chudackova, K. (2004). Changes of properties of polymer packaging
films during high pressure treatment. J Food Eng. 61: 545–549.
Donsi, F., Giovanna, F., and Gianpiero, P. (2010). Applications of pulsed electric field treatments for the
enhancement of mass transfer from vegetable tissue. Food Eng Rev. 2: 109–130.
Dunn, J.E., Clark, R.W., Asmus, J.F., Pearlman, J.S., Boyer, K., Painchaud, F., and Hofmann, G.A. (1989).
Methods for Preservation of Foodstuffs. San Diego, CA: Maxwell Laboratories Inc. US Patent
4871559.
Dunn, J., Ott, T., and Clark, W. (1995). Pulsed-light treatment of food and packaging. Food Technol. 49(9): 95–98.
Dury-Brun, C.C., Chalier, P., Desorby, S.P., and Voilley, A.E. (2007). Multiple mass transfers of small volatile
molecules through flexible food packaging. Food Reviews Internat. 23(3): 199–255.
Eie, T. (2009). Light protection from packaging. In: Encyclopedia of Packaging Technology, pp. 655–659.
Edited by K.L. Yam. Hoboken, NJ: John Wiley & Sons.
El Makhzoumi, Z. (1994). Effect of irradiation of polymeric packaging material on the formation of volatile
compounds. In: Food Packaging and Preservation, pp. 88–99. Edited by M. Mathlouthis. London:
Blackie Academic & Professional.
Elmnasser, N., Guillou, S., Leroi, F., Orange, N., Bakhrouf, A., and Federighi, M. (2007). Pulsed light system
as a novel food decontamination technology: A review. Canadian J Microbial. 53: 813–821.
Esbelin, J., Mallea, S., Ram, A.F.J.R., and Carlin, F. (2013). Role of pigmentation in protecting Aspergillus
niger conidiospores against pulsed light radiation. Photochem Photobiol. 89: 758–761.
Evrendilek, G.A., Dantzer, W.R., Streaker, C.B., Ratanatriwong, P. and Zhang, Q.H., (2001). Shelf-life evalu-
ations of liquid foods treated by pilot plant pulsed electric field system. J Food Process Preserv. 25(4):
283–297.
Evrendilek, G.A., and Zhang, Q.H. (2003). Effects of pH, temperature and pre-PEF treatments on PEF and
heat inactivation of Escherichia coli O157: H7. J Food Protect. 66: 755–759.
FDA/CFSAN. (2005). Kinetics of microbial inactivation for alternative food processing technologies: execu-
tive summary. US Food and Drug Administration. Center for Food Safety and Applied Nutrition.
Farkas, D.F., and Hoover, D.G. (2000). High pressure processing. J Food Sci. 65: 47–64.
Food Safety. www.sterigenics.com (2005).
Franz, C.M.A.P., Specht, I., Cho, G.S., Graef, V., and Stahl, M.R. (2009). UV-C inactivation of microorganisms
in naturally cloudy apple juice using novel inactivation equipment based on dean vortex technology. Food
Cont. 20: 1103–1107.
Galotta, M.J., Ulloa, P.A., Guarda, A., Gavara, R., and Miltz, J. (2009). Effect of high-pressure food process-
ing on the physical properties of synthetic and biopolymer films. J Food Sci. E304–E311.
Garcia, A.F., Butz, P., Bognar, A., and Tauscher, B. (2001). Antioxidative capacity, nutrient content and sen-
sory quality of orange juice and an orange_lemon_carrot juice product after high pressure treatment and
storage in different packaging. Eur Food Res Technol. 213: 290–296.
Goetz, J., and Weisser, H. (2002). Permeation of aroma compounds through plastic films under high pressure:
in-situ measuring method. Innov Food Sci Emerging Technol. 3: 25–31.
Gómez-López, V.M., Devlieghere, F., Bonduelle, V., and Debevere, J. (2005a). Intense light pulses decontami-
nation of minimally processed vegetables and their shelf-life. Int J Food Microbiol. 103: 79–89.
Gómez-López, V.M., Ragaert, P., Debevere, J., and Devlieghere, F. (2007). Pulsed light for food decontamina-
tion: A review. Trends Food Sci Technol. 18: 464–473.
Goodner, J.K., Braddock, R.J., Parish, M.E., and Sims, C.A. (1999). Cloud stabilization of orange juice by high
pressure processing. J Food Sci. 64 (4): 699–700.
Goulas, A.E., Riganakos, K.A., Badeka, A., and Kontominas, M.G. (2002). Effect of ionizing radiation on
the physicochemical and mechanical properties of commercial monolayer flexible plastics packaging
materials. Food Additives Contam. 19: 1190–1199.
Grahl, T., and Markl, H. (1996). Killing of microorganisms by pulsed electric fields. Applied Microbiol
Biotechnol. 45: 148–157.
Grecz, N., Rowley, D.B., and Matsuyama, A. (1983). The action of radiation on bacteria and viruses.
In: Preservation of Foods by Ionizing Radiation, pp. 167–218. Edited by E.S. Josephson and M.S.
Petersons. Boca Raton, FL: CRC Press.
Guillard, V., Mauricio-Iglesias, M., and Gontard, N. (2010). Effect of novel food processing methods on
packaging: Structure, composition, and migration properties. Critical Review Food Sci Nutrit. 50(10):
969–988.
Han, J.H. (2007). Packaging for non-thermally processed foods. In: Packaging for Non Thermal Processing
of Food, p. 13. Ames, IA: Blackwell Publishing.
Han J., Gomes-Feitosa, C.L., Castell-Perez, E., Moreira, R.G., and Silva, P.F. (2004). Quality of packaged
romaine lettuce hearts exposed to low-dose electron beam irradiation. Lebensmittel Wissenschaft
Technologie. 37: 705–715.
Heinrich, V., Zunobovic, M., Bergmair, J., Kneifel, W., and Jager, H. (2015). Post packaging application of pulsed
light for microbial decontamination of solid foods. Innovat Food Sci Emerging Technol. 30: 145–156.
Hernandez-Andres, A., Gomez-Guillen, C., Montero, P., Perez-Mateos, M. (2005). Partial characterization of
protease activity in squid (Todaropsis eblanae) mantle: Modification by high-pressure treatment. J Food
Sci. 70(4): C239–C245.
Hodgins, A.M., and Mittal, G.S. (2003). Pulsed electric field assisted non-thermal pasteurization of pumpable
foods. Food Science Central, Food Info Online Features 2 July 2003, IFIS Publ.
Hog, G., and Cumberbatch, G.M. (1991). Stability of Surlyn ionomer films to ionizing radiation. J Plastic Film
Sheeting. 7: 221–246.
Hogan, E., Kelly, A.L., and Sun, W.D. (2005). High pressure processing of foods. In: An Overview in Emerging
Technologies for Food Processing, pp. 3–12. London, UK: Elsevier Academic Press.
Hulsheger, H., and Niemann, E.G. (1980). Lethal effects of high voltage pulses on E. coli K12. Radiation
Environ Biophy. 18: 281–288.
Jin, Z.T., and Zhang, Q.H. (2002). Cost evaluation of a commercial scale PEF system. In: IFT Annual Meeting,
June 15–19, 2002. Anaheim, CA: Institute of Food Technologists, (p. 229/91E-21).
Jo, C., Jin, S.K., and Ahn, D.U. (2000). Colour changes in irradiated cooked pork sausage with different fat
sources and packaging during storage. Meat Sci. 55(1): 107–113.
Katz, F. (1999). Smart packaging adds a dimension to safety. Food Technol. 53(11): 106.
Keklik, N.M., Demirci, A., and Puri, V.M. (2009). Inactivation of Listeria monocytogenes on unpackaged and
vacuum-packaged chicken frankfurters using pulsed UV-light. J Food Sci. 74(8): M431–M439.
Keklik, N.M., Demirci, A., and Puri, V.M. (2010). Decontamination of unpackaged and vacuum-packaged
boneless chicken breast with pulsed ultraviolet light. Poultry Sci. 89: 570–581.
Killoran, J.J. (1983). Packaging irradiated food. In: Preservation of Food by Ionizing Radiation, pp. 317–326.
Edited by E. Josephson and M. Peterson. Boca Raton, FL: CRC Press, vol. 2.
Kim, Y.H., Nam, K.C., and Ahn, D.U. (2002). Irradiation of packaged food. Meat Sci. 61(3): 257–265.
Kinosita, K., and Tsong, T.Y. (1977). Formation and resealing of pores of controlled sizes in human erythro-
cyte membrane. Nature. 268: 438–440.
Kirwan, M.J., and Strawbridge, J.W. (2003). Plastics in food packaging. In: Food Packaging Technology,
pp. 174–240. Edited by R. Coles, D. McDowell, and M.J. Kirwan. Oxford: CRC Press.
Komolprasert, V. (2016). Packaging food for radiation processing. Radiation Phy Chem. 129: 35–38.
Komolprasert, V., McNeal, T.P., Agrawal, A., Adhikari, C., and Thayer, D.W. (2001). Volatile and nonvolatile
compounds in irradiated semi-rigid crystalline poly (ethylene terephthalate) polymers. Food Additives
Contaminan. 18(1): 89–101.
Komolprasert, V., and Morehouse K.M. (2004). Irradiation of Food and Packaging Recent developments, ACS
Symposium Series 875. Washington, DC: American Chemical Society.
Krishnamurthy, K., Demirci, A., Puri, V.M., and Cutter, C.N.N. (2004). Effect of packaging materials on inacti-
vation of pathogenic microorganisms on meat during irradiation. Transactions of the ASAE, 47: 1141–1149.
Kubel, J., Ludwig, H., Marx, H., and Tauscher, B. (1996). Diffusion of aroma compounds into packaging films
under high pressure. Packag Technol Sci. 9: 143–152.
Kumar, P., Han, J.H. (2012). Packaging materials from non-thermal processing of foods and beverages.
In: Emerging Food Packaging Technologies, pp. 323–334. Edited by K. Yam and D.S. Lee. Oxford,
UK: Woodhead Publishing.
Lacorix, M. (2014). Irradiation in emerging technologies for food processing. Edited by W.D. Sun. London,
UK: Academic Press Elsevier, 2nd edition, pp. 293–295.
Lacroix, M., Marcotte, M., and Ramaswamy, H. (2002). Irradiation in combination with others processes
for the preservation of fruits, vegetables, nuts and spices. In: Handbook of Postharvest Technology,
pp. 623–652. New York: Marcel Dekker.
Lagunas-Solar, M.C., and Gómez-López, V.M. (2006). COST Sub-committee report on UV units, UV sources
specifications and experimental procedures.
Lambert, Y., Demazeau, G., Largeteau, A., and Bouvier, J.M. (2000). Packaging for highpressure treatments
in the food industry. Packag Technol Sci. 13: 63–71.
Le-Bail, A., Hamadami, N., and Bahuaud, S. (2006). Effect of high-pressure processing on the mechanical
and barrier properties of selected packagings. Packag Technol Sci. 19: 237–243.
Lee, J.H., Sung, T.H., Lee, K.T., and Kim, M.R. (2004). Effect of gamma-irradiation on colour, pungency, and
volatiles of Korean red pepper powder. J Food Sci. 69: C585–C592.
Levy, C., Aubert, X., Bornard, I., and Carlin, F. (2012). Relevant factors affecting microbial surface decon-
tamination by pulsed light. Internat J Food Microbiol. 152: 168–174.
Li, B., and Logan, B.E. (2005). The impact of ultraviolet light on bacterial adhesion to glass and metal
oxide-coated surface. Colloids and Surfaces B: Biointerfaces. 41(2): 153–161.
Linton, M., and Patterson, M.F. (2000). High pressure processing of foods for microbiological safety and qual-
ity. Acta Microbiol Immunol Hung. 47(2–3): 175–182.
Lopez-Gonzalez, V., Murano, P.S., Brennan, R.E., and Murano, E.A. (2000). Sensory evaluation of ground
beef patties irradiated by gamma rays versus electron beam under various packaging conditions. J Food
Qual. 23: 195–204.
Mafu, A.A., Roy, D., Goulet, J., and Savoie, L. (1991). Characterization of physicochemical forces involved in
adhesion of Listeria monocytogenes to surfaces. Applied Environ Microbiol. 57(7): 1969–1973.
Marque, D., Feigenbaum, A., and Riquet, A.M. (1995). Consequences of polypropylene film ionisation on the
food/packaging interactions. J Poly Eng. 15: 101–115.
Masuda, M., Saito, Y., Iwanami, T., and Hirai, Y. (1992). Effects of hydrostatic pressure on packaging mate-
rials for food. In: High Pressure and Biotechnology, pp. 545–547. Edited by C. Balny, R. Hayashi,
K. Heremans, and P. Massons. London: John Libbey Eurotext.
Matsui, T., Inoue, M., Shimoda, M., and Osajama, Y. (1991). Sorption of volatile compounds into electron
beam irradiated EVA film in the vapour phase. J Sci Food Agri. 54: 127–135.
Meyer, R., Cooper, K.L., Knorr, D., and Lelieveld, H.L.M. (2000). High pressure sterilisation of foods. Food
Technol. 54(11): 67–72.
Min, C.S., Zhang, Q.H., and Han, H.J. (2014). Packaging for non thermal food processing. In: Innovation in Food
Packaging, pp. 516–531. Edited by Han, H.J. Boston, MA: Academic Press, Elsevier, 2nd edition, p. 589.
Min, S., Jin, Z.T., and Zhang, Q.H. (2003a). Commercial scale pulsed electric field processing of tomato juice.
J Agri Food Chem. 51: 3338–3344.
Min, S., Jin, Z.T., Min, S.K., Yeom, H. and Zhang, Q.H. (2003b). Commercial scale pulsed electric field pro-
cessing of orange juice. J Food Sci. 68(4): 1265–1271.
Min, S., and Zhang, Q.H. (2007). Packaging for high pressure processing, irradiation and pulsed electric field
processing. In: Packaging for Non Thermal Processing of Food, pp. 70–71. Edited by H.J. Han. Ames,
IA: Blackwell Publishing. 240p.
Morris, C., Brody, A.L., and Wicker, L. (2006). Non-thermal food processing/preservation technologies: A
review with packaging implications. Packag Technol Sci. 20: 275–286.
Mortensen, G., Bertelsen, G., Mortensen, B.K., and Stapelfeldt, H. (2004). Light-induced changes in packaged
cheeses: A review. Internat Dairy J. 14(85), 102.
Moura, E.A.B., Ortiz, A.V., Wiebeck, H., Paula, A.B.A., Silva, A.L.A., and Silva L.G.A. (2004). Effects of
gamma radiation on commercial food packaging films: Study of changes in UV/VIS spectra. Radiat Phy
Chem. 71(1–2): 201–204.
Murano, E.A., Murano, P.S., Brennan, R.E., Shenoy, K., and Moreira, R.G. (1999). Application of high hydro-
static pressure to eliminate Listeria monocytogenes from fresh pork sausage. J Food Protect. 62(5):
480–483.
Muredzi, P. (2012). Emerging Non-Thermal Food Processing Technologies. North Andover, MA: Cambridge
Brick House.
Nam, K.C., and Ahn, D.U. (2002). Mechanisms of pink colour formation in irradiated precooked turkey breast
meat. J Food Sci. 67(2): 600–607.
Norton, T., Delgado, A., Hogan, E., Grace, P., and Sun, D.W. (2008). Simulation of high pressure freezing
processes by enthalpy method. J Food Eng. 91: 260–268.
NOVA Institute. (2013). Market study on bio-based polymers in the world. Capacities, production and applica-
tions: Status quo and trends towards 2020.
Odriozola-Serrano, I., Aguiló-Aguayo, I., Soliva-Fortuny, R., and Martín-Belloso, O. (2013). Pulsed electric
fields processing effects on quality and health-related constituents of plant-based foods. Trends Food
Olson, D.G. (1998). Irradiation of Food. Food Technol. 52: 56–62.
Oms-Oliu, G., Martin-Belloso, O., and Soliva-Fortuny, R. (2010). Pulsed light treatments for food preserva-
tion: A review. Food Bioprocess Technol. 3: 13–23.
Ozen, B.F., and Floros, J.D. (2001). Effects of emerging food processing techniques on the packaging materi-
als. Trends Food Sci Technol. 12: 60–67.
Food Processing, pp. 279–306. London: Elsevier Academic Press, 762p.
Palou, E., Hernandez-Salgado, C., Lopez-Malo, A., Barbosa-Canovas, G.V., Swanson, B.G., and Welti-Chanes, J.W.
(2000). High pressure-processed guacamole. Innov Food Sci Emerg Technol. 1: 69–75.
Pandrangi, S., and Balasubramaniam, V.M. (2005). High pressure processing of salads and ready meals.
In: Emerging Technologies for Food Processing, pp. 33–43. Edited by Da-Wen Sun, London, UK:
Elsevier Academic Press, 762p.
Pentimalli, M., Ragni, P., Righini, G., and Capitani, D. (2000). Polymers and paper as packaging materials of
irradiated food: An NMR study. Radiation Phy Chem. 57(36): 385–388.
Pilette, L. (2003). Effects of ionizing treatments on packaging: Food simulant combinations. Packaging pump-
able foods. In: Food Science Central, Food Info Online Features 2 July, IFIS Publ.
Pospíšil, J., and Nešpůrek, S. (2008). Polymer additives. In: Plastic Packaging, pp. 63–68. Edited by O.G.
Piringer and A.L. Baner. Weinheim Germany: Wiley-VCH.
Qin, B.L., Pothakamury, U.R., Vega, H., Martin, O., Barbosa-Canovas, G.V., and Swanson, B.G. (1995). Food
pasteurization using high intensity pulsed electric fields. Food Technol. 49: 55–60.
Qiu, X., Sharma, S., Tuhela, L., Jia, M., Zhang, Q.H. (1998). An integrated PEF pilot plant for continuous non-
thermal pasteurization of fresh orange juice. Trans Am Soc Agric Eng. 41(4): 1069–1074.
Quass, D.W. (1997). Pulsed electric field processing in the food industry. In: A Status Report on Pulsed Electric
Field, p. 2335. Palo Alto, CA: Electric Power Research Institute.
Raab, M., Kotulák, L., Kolařík, J., and Pospíšil, J. (1982). The effect of ultraviolet light on the mechanical
properties of polyethylene and polypropylene films. J Applied Polymer Sci. 27(7): 2457–2466.
Radiation Applications Research Branch, Canada, www.iaea.org/icgfi/packaging-dat-canada.pdf (2002).
Raghubeer, E.V., Dunne, C.P., Farkas, D.F., and Ting, E.Y. (2000). Evaluation of batch and semicontinuous appli-
cation of high hydrostatic pressure on foodborne pathogens in salsa. J Food Protect. 63(12): 1713–1718.
Rajkovic, A., Tomasevic, I., Smigic, N., Uyttendaele, M., Radovanovic, R., and Devlieghere, F. (2010). Pulsed
UV light as an intervention strategy against Listeria monocytogenes and Escherichia coli O157:H7 on
the surface of a meat slicing knife. J Food Eng. 100: 446–451.
Raso, J., Calderon, M.L., Gongora, M., Barbosa-Canovas, G.V., and Swanson, B.G. (1998). Inactivation of
mold ascospores and conidio spores suspended in fruit juices by pulsed electric fields. Lebensmittel
Wissenschaft Technologie. 31: 668–672.
Ringus, D. L., and Moraru, C.I. (2013). Pulsed light inactivation of Listeria innocua on food packaging mate-
rials of different surface roughness and reflectivity. J Food Eng. 114(3): 331–337.
Robert, F.S., Mariona, P.V., Olga, B.M., and Pedro, M.E. (2017). Effect of pulsed electric field on the anti-
oxidant potential of apples stored at different temperature. Postharvest Biol Technol. (article in press)
informa http://dx.doi.org/10.1016/j.postharvbio.2017.03.015
Rojas De Gante, C., and Pascat, B. (1990). Effects of p-ionizing radiation on the properties of flexible packag-
ing materials. Packag Technol Sci. 3: 97–105.
Saldaña, G., Álvarez, I., Condón, S., and Raso, J. (2014). Microbiological aspects related to the feasibility of
PEF technology for food pasteurization. Crit Rev Food Sci Nutr. 54: 1415–1426.
Shauwecker, A., Balasubramaniam, V.M., Sadler, G., Pascall, M.A., and Adhikari, C. (2002). Influence of high-
pressure processing on selected polymeric materials and on the migration of a pressure-transmitting
fluid. Packag Technol Sci. 15: 255–262.
Smelt, J.P. (1998). Recent advances in the microbiology of high pressure processing. Trends Food Sci Technol.
9: 152e158.
Smith, J.S., and Pillai, S. (2004). Irradiation and food safety. Food Technol. 58: 48–55.
Sohn, K.H. and Lee, H.J. (1998). Effects of high pressure treatment on the quality and storage of kimchi. Int
J Food Sci Technol. 33(4): 359–365.
Sommers, C.H., Cooke, P.H., Fan, X., and Sites, J.E. (2009). Ultraviolet light (254 nm) inactivation of Listeria
monocytogenes on frankfurters that contain potassium lactate and sodium diacetate. J Food Sci. 74(3):
M114–M119.
Takahashi, F., Pehrsson, P.E., Rovere, P., and Squarcina, N. (1998). High-pressure processing of fresh orange
juice. Ind Conserv. 73(4): 363–368.
Takeshita, K., Shibato, J., Sameshima, T., Fukunaga, S., Isobe, S., Arihara, K., et al. (2003). Damage of yeast
cells induced by pulsed light irradiation. Internat J Food Microbiol. 85: 151–158.
Tao, Y., Sun, D.S., Hogan, E., and Kelly, A.L. (2007). High-pressure processing of foods. In: An Overview in
Emerging Technologies for Food Processings, pp. 1–5. Edited by W.D. Sun. London, UK: Academic
Press, Elsevier.
Thakur, B.R., Singh, R.K., and Nelson, P.E. (1996). Quality attributes of processed tomato products: A review.
Food Rev Internat. 12: 375–401.
Thayer, D. (1988). Chemical change in food packaging resulting from ionizing radiation. ACS Symp Ser 365:
181–194.
Toepfl, S., Heinz, V. and Knorr, D. et al. (2005). Overview of pulsed electrical field processing for food. In
Emerging Technologies of Food Processing, pp. 69–87. Edited by Da Wen Sun. Elsevier.
Tuboly, E., Lebovics, V.K., Gaal, O., Meszaros, L., and Farkas, J. (2003). Microbiological and lipid oxidation
studies on mechanically deboned turkey meat treated by high hydrostatic pressure. J Food Eng. 56(2/3):
241–244.
USFDA, Code of Federal Regulations 21, parts 170–199 (1987), also 7405 (February 1989).
Vanderroost, M., Ragaert, P., Devlieghere, F., and De Meulenaer, B. (2014). Intelligent food packaging: The
next generation. Trends Food Sci Technol. 39: 47–62.
Vega-Mercado, H., Pothakamury, U.R., Chang, F.J., Barbosa-Canovas, G.V., and Swanson, B.G. (1996).
Inactivation of Escherichia coli by combining pH, ionic strength and pulsed electric fields hurdles.
Food Res Internat. 29: 117–121.
Wekhof, A. (2000). Disinfection with flash lamps. J Pharma Sci Technol. 54(3): 264–276.
Wekhof, A., and Trompeter, F.J. (2001). Pulsed UV disintegration (PUVD): A new sterilisation mechanism
for packaging and broad medical–hospital applications. In: The First International Conference on
Ultraviolet Technologies. June 14–16, 2001, Washington, DC.
Woodling, S.E., and Moraru, C.I. (2005). Influence of surface topography on the effectiveness of pulsed
light treatment for the inactivation of Listeria innocua on stainless steel surfaces. J Food Sci. 70(7):
M345–M351.
World Health Organization (WHO). (1999). High dose irradiation: Wholesomeness of foodirradiated with
doses above 10 kGy. Report of a Joint Food and Agriculture Organization (FAO), International Atomic
Energy Agency (IAEA), World Health Organization (WHO) Study Group.
Wouters, P.C., Bos, A.P., and Ueckert, J. (2001). Membrane permeabilization in relation to inactivation kinet-
ics of Lactobacillus species due to pulsed electric fields. Applied Environ Microbiol. 67: 3092–3101.
Yeom, H.W., Charles, B., Streaker, Q., Zhang, H., and Min, D.B. (2000). Effect of pulsed electric fields on the
quality of orange juice and comparison with heat pasteurization. J Agric Food Chem. 48(10): 4597–4605.
Young, A.L. (2003). Food irradiation. Environ Sci Pollution Res. 10: 82–88.
Zhang, Q., Barbosa-Canovas, G.V., and Swanson, B.G. (1995a). Engineering aspects of pulsed electric field
pasteurization. J Food Eng. 25: 261–281.
Zhang, Q., Monsalve-Gonzalez, A., Qin, B.L., Barbosa-Canovas, G.V., and Wanson, B.G. (1994). Inactivation
of Saccharomyces cerevisiae in apple juice by square-wave and exponential decay pulsed electric fields.
J Food Process Eng. 17: 469–478.
Zhang, Q., Qin, B.L., Barbosa-Canovas, G.V., and Swanson, B.G. (1995b). Inactivation of E. coli for food
pasteurization by high-strength pulsed electric fields. J Food Process Preservat. 19: 103–118.
Ziegler, R., Schanderl, S.H., and Markakis, P. (1968). Gamma irradiation and enriched CO2 atmosphere stor-
age effects on the light-induced greening of potatoes. J Food Sci. 33(5): 533–535.
ChaptEr 20
Commercialization and regulatory Issues

of Non-thermal processed Foods
Feby Luckose, B. S. Mamatha, and O. P. Chauhan
CONtENtS
20.1 Introduction ......................................................................................................................... 417

20.2 Nutritional Aspects of Non-thermally Processed Food ...................................................... 418
20.3 Energy Requirements and Environmental Concerns .......................................................... 420
20.4 Packaging Issues ................................................................................................................. 422
20.5 Consumer Response ............................................................................................................ 423
20.6 Regulatory Aspects ............................................................................................................. 424
20.7 Current Status of Commercial Applications ....................................................................... 426
20.8 Conclusion .......................................................................................................................... 428
References ...................................................................................................................................... 428
20.1 INtrODUCtION
Food-borne illnesses caused by bacterial pathogens have raised concern about the safety of
food over the past few decades. As a result, testing for biological hazards in foods is increasing
significantly as awareness of food-borne diseases from microbial pathogens and their toxins gains
more public attention and a higher regulatory profile. In addition, with hazard analysis and critical
control point (HACCP) systems in place in many food industries, risk-based food safety assess-
ment has become mandatory. Although the food industry, regulators and consumers would favour a
totally risk-free food supply, zero-risk is neither practical nor achievable. Development of methods
for identifying health hazards and predicting food safety is of high priority.
Increased knowledge about food and health among the general public has increased the
demand for minimally processed food with high nutritional and sensory profile (Chipurura and
Muchuweti, 2010). One of the common methods for food preservation is by thermal treatment or
elevated temperature treatments [sterilization by ultra-high temperature (>100°C), pasteurization
(<100°C), and cooking (100°C)]. Thermal treatments have an irreversible effect on nutritive and
sensory parameters of the food. Therefore, preservation by non-thermal treatment is gaining a lot
of demand in the present market and have become an alternative method for thermal preservation
(Guerrero-Beltran and Barbosa-Canovas, 2005). Retention of flavour and aroma after a processing
step is a huge advantage for Asian foods, which often have stronger taste and aroma compared to
Western foods. The vast variety of flavourful food in Southeast Asia such as fruits and fruit juices,
sauces and seasonings, cereals, grains, flour and starches, seafood and meat products, snacks, and
417
traditionally preserved foods (fermented, salted, added sugar) creates potential for the commercial
application of non-thermal processing technologies for shelf life extension of food.
The term “non-thermal processing” is used to designate technologies that are effective at ambient
or sub-lethal temperatures. Irradiation, high-pressure processing, pulsed electric fields, ultrasound,
pulsed light (PL), ozone and ultraviolet light are some non-thermal technologies that can inactivate
micro-organisms without the use of high temperature (Pereira and Vicente, 2010).
Food preservation by heat is achieved by microbial inactivation. This is brought about by
alteration in cellular structures like DNA strand breakage, cell membrane rupture or by altering
cell functions like membrane permeability in micro-organisms (Lado and Yousef, 2002). Limited
information is available about the mechanisms of inactivation of microorganisms by alternative
preservation technologies. To speed up the implementation of these technologies, a huge number
of studies has been undertaken by food researcher all over the world to decipher the mechanisms
of cell inactivation by non-thermal means. Nevertheless, these new and alternative food processing
methods, as well as novel combinations of existing methods, are continually being sought by
industry in an effort to produce better quality food.
Manufacturing, processing, and packaging of modern processed foods and traditional foods
have become innovative and modernized to an incomparable extent. These novel technologies
have scope for producing high-quality and safe food products but their current limitations like
high investment costs, full control of variables associated with the process operation, lack of
regulatory approval and, importantly, consumer acceptance has delayed implementation of these
technologies to their full potential at a commercial scale. Permission from the governmental
regulatory agencies to use the modern technology is one of the key requirements for the
commercialization of new thermal processes (Han, 2008). There are not much scientific data to
address the problems faced in commercialization. The efficacy of alternate methods over heat
processing in improving the nutritional profile of food as well as certain issues related to nutritional
aspects that needs to be addressed have been briefly discussed in the first session, followed
by sections on packaging concerns, energy requirements and environmental impact and most
importantly response of consumers for non thermally processed food. There is relative shortage
of published information in these areas of novel alternate technologies. Finally, the regulatory
aspects of implementing these new processes and the current status of their commercialization
have been discussed.
20.2 NUtrItIONaL aSpECtS OF NON-thErMaLLY prOCESSED FOOD
Novel, non-thermal technologies promise to treat foods without destroying the nutritional com-
ponents and sensorial characteristics that are normally affected during heat treatment. The latest
techniques today applied in both research institutes and food industries are to shorten processing
times, control Maillard reactions, improve products quality, and enhance functionality. These tech-
nologies are of specific interest to the food industry, because they not only provide attractive alter-
natives to conventional methods (thermal processing), which often produce undesirable changes in
foods, affecting the balance between high quality and safety, but also create new ingredients and
products because of the specific actions on biological materials and food constituents (Knorr et al.,
2002). Product safety (chemical safety and microbiological safety) is the precondition for a food to be
in the market. The general advantage of these advanced technologies is the elimination of chemical
additives. With diet and health firmly established as a long-term lifestyle, it has become important
to understand the impact of these innovative technologies on the nutritional value of foods.
Literature studies indicate that high pressure processing (HPP) preserves the nutritional value
in food products. Yen and Lin (1996) suggested that ascorbic acid degradation after high hydrostatic
pressure (HHP) processing could take place during storage and it could be eliminated by lowering
CoMMerCiALiZAtion AnD reGuLAtory issues oF non-tHerMAL ProCesseD FooDs 419
storage temperature. Furthermore, it has been reported that different HHP combinations had different
influences on the stability of vitamin C in guava puree during storage. High-pressure treatments
modified the mechanism of anthocyanin degradation by affecting the molecules involved in the
kinetics of reaction, such as enzymes. It is obvious that HPP treatment influences the phytochemical
stability and the extraction yield of bioactive compounds (Prasad et al., 2009). Excessive heating
typically causes off-flavour production and other adverse quality changes. Unlike during thermal
pasteurization, the product temperature during HPP or PEF is increased only slightly. Also, high
pressure does not affect covalent bonds, and development of flavours alien to the products is therefore
prevented, hence maintaining the natural qualities of the products (Stute et al., 1996).
Irradiation of food products causes minimal modification in the flavour, colour, nutrients, taste,
and other quality attributes of food (Alothman et al., 2009b). However, the levels of modification (in
flavour, colour nutrients, taste etc.) might vary depending on the basic raw material used, irradiation
dose delivered, and on the type of radiation source employed (gamma, X-ray, UV, electron beam)
(Bhat et al., 2007; Bhat and Sridhar, 2008). Irradiation induces negligible or subtle losses of bioactive
compounds as it does not substantially raise the temperature of food during processing (Wood
and Bruhn, 2000). No major changes in the carotenoids content were recorded (Alighourchi et al.,
2008). The decrease in antioxidant compounds is attributed to the formation of radiation-induced
degradation products or the formation of free radicals (Sajilata and Singhal, 2006; Wong and Kitts,
2001). A different study showed total phenol and flavonoid content increases in guava and banana on
UV treatment (Alothman et al., 2009a, 2009b). Cheng et al. (2007) reported an increase in ascorbic
content of guava juice during sonication (from 110 to 119 mg/100 mL) and a significant increase
in combination of sonication and carbonation (125 mg/100 mL) which might be due to cavitation
effects caused by carbonation and sonication.
PEF treatment only to a slight extent modify the phenolic compounds, vitamin C and antioxidant
contents (Morales-de la Peña et al., 2010a, 2010b) compared to conventional thermal treatment.
In PEF treatment, an important nutritional concern is the interaction between electrode materials
and foods that still needs to be understood. Systematic work on safety issues (e.g., on free-radical
production) should be carried out to assure regulatory approval.
Ozone has been successfully implemented as a surface decontamination technology. However,
from the nutritional point of view it may not be a desirable treatment for certain products like fruit
juices. Studies revealed that ozone is expected to cause the loss of antioxidant compounds, because
of its strong oxidizing activity (Barboni et al., 2010). It is seen that ozone treatment significantly
decreases the vitamin C content in fruits (Alothman et al., 2009a, 2009b). The main factor that
contributes to the degradation of ascorbic acid in ozone-treated fruit is the activation of ascorbate
oxidase. This enzyme is activated under stress conditions, such as chemical exposure. Ascorbate
oxidase has been reported to promote the degradation of ascorbic acid to dehydroascorbic acid (Lee
and Kader, 2000). Studies show that the effects of ozone on physiology and quality of fruits vary
according to chemical composition of food, ozone dose, and application type and time (Whangchai
et al., 2006; Karaca and Velioglu, 2007; Cullen et al., 2010).
Several studies report that technologies such as high hydrostatic pressure and pulsed electric
fields could increase the extraction of bioactive compounds from fruit and vegetables and increase
their healthy potential, probably improving their bioavailability. The research studies carried out on
bioavailability and antioxidant properties of the non-thermal processed vegetable products as orange
juice and vegetable soup, indicate high vitamin C bioavailability and improved health potential by
an increase in plasma vitamin C and a decrease of the oxidative stress and inflammation biomarkers
in healthy humans. From the point of view of commercialization, industries could provide to the
consumers new healthy vegetable-derived products by combining these technologies with other
food processing treatments or by using them to extract bioactive ingredients from low-cost raw
material or vegetable by-products, to obtain functionalized products with better sensorial quality
and assured safety.
Further studies on kinetic data associated with the impact of non thermal technologies on bioac-
tive components in various food systems are needed. Nutritional aspects of these non-thermally-
treated foods have been neglected and more work is still required to be carried out in this area.
More information on interactions between these processes and toxins or allergens is also needed. It
is also necessary to obtain healthy minimal processed foods and to design functional foods to get
better consumer quality of life (Knorr et al., 2002). Finally, it is worth noting that pressure-induced
phase transitions such as crystallization of lipids or thawing or freezing of high-moisture systems
offer numerous opportunities for process or product development. Employing non-thermal methods
to synthesize bioactive compounds in foods is of great nutritional benefit which provides attractive
opportunities for the food industry (Sanchez-Moreno et al., 2009).
20.3 ENErGY rEQUIrEMENtS aND ENVIrONMENtaL CONCErNS
A food process operation system involves addition or removal of thermal energy to ensure
microbiologically safe, improved quality, and enhance the product shelf-life. The food industry
ranks 4th in energy use after the chemical, mining, and paper industries; depending on the product,
food processing can account for up to one-third of energy use in the food sector due to consumer
demand for convenience, indulgence, and fast foods (Hendrickson, 1996). Among traditional pres-
ervation methods, canning, freezing, and drying are considered the most energy-intensive process-
ing operations (Hendrickson, 1996). Alternative food preservation technologies include changes
in heating methods that can reduce energy consumption. The food industry continues to develop
solutions toward sustainable food manufacturing and more efficient energy use. While conventional
processes (high hydrostatic pressure, PEF, high-intensity ultrasound, UV light, PL, ionizing radia-
tion, and oscillating magnetic fields) primarily utilize thermal energy, advanced processes utilize
mechanical, electromagnetic, light, electrical, and other forms of energy to accelerate reactions,
such as inactivation of microorganisms (Butz and Tauscher, 2002). Some of these treatments may
generate internal energy (e.g., adiabatic heating and resistive heating during HHP and PEF, respec-
tively); however, they are classified as non-thermal once, contrarily to thermal processing technolo-
gies, because they eliminate the use of elevated temperatures to kill the microorganisms, avoiding
the harmful effects of heat on flavour, colour, and nutritive value of foods.
PEF technology advocates less energy can be expended to achieve the same operational objectives
in cell disintegration and microbial inactivation (Toepfl et al., 2006). In a study, three levels of
energy evaluation for each technology including internal energy, applied energy, and consumed
energy were reviewed. The comparison of the specific energy for the different technologies on
the inactivation of Escherichia coli in apple juice was explored. Analysis of energy consumption
of these technologies concluded that UV consume less specific energy than HTST, PEF, and HPP
(Rodriguez-Gonzalez et al., 2015). The differences in energy consumption by different technologies
demonstrate that there is potential of improvement in most technologies. Loss in energy occur at
the time of transmission and conversion of energy into different forms, such as from electrical to
thermal energy, depending on the process and specific equipment design (technology does have
different energy conversion efficiency) (Rodriguez‐Gonzalez et al., 2015). However, there are no
reports available on a comparative analysis of the fundamentals of energy conversation, evaluation
approaches and energy usage of alternative technologies for specific unit operations.
PL systems have comparatively low operation costs and do not contribute significantly negative
to the environmental impact, because they have the potential to eliminate microorganisms
without the need for any chemicals. Further, they do not produce volatile organic compounds and
generate only reduced amounts of solids wastes. However, the poor penetrating power of light
(requires transparency and surface smoothness of the product to be treated) and high investment
costs (€ 300,000–800,000) are currently limiting PL applications (Pereira and Vicente, 2010).
In PEF treatment, theoretically, the temperature increase of the treated food can be calculated from
the effective pulse energy. Because of this, direct temperature measurements can also be used as an
indirect way to estimate energy use and may illustrate potential energy losses in the system. On the
other hand, PEF and HPP treatments showed smaller differences in specific energy when compared
to HTST. For example, PEF requires energy between 70 and 41 J/g, HPP between 483 and 338 J/g,
and HTST between 228 and 167 J/g. PEF has the potential to lower the energy consumption by
optimizing pulsing conditions (changing to square instead of exponential pulses and by frequency
and flow optimization) and increasing processing temperatures. Based on pressure rise HPP
energy is estimated and thus, there is a chance of improvement except by increasing the process
temperature. Use of semi-continuous systems, or better utilization of pressure vessel volume by
modifying packaging design, are potential approaches that can improve energy utilization within a
pressure vessel.
Traditional, thermal-based food processes are often well optimized to recover a significant part
of the energy in heat exchangers where the food’s enthalpy is used to heat or cool. Due to the emerg-
ing nature of alternative food preservation technologies, very limited efforts have been made to
develop such energy recovery approaches. However, it is evident from the literature that there is
a great potential to improve equipment and system and process design from an energy efficiency
and recovery perspective. For example, the design of high-pressure intensifiers can be optimized
to minimize energy loss during conversion of electrical energy into mechanical energy. Similarly,
during PEF processing a portion of dissipated electrical energy can be recovered as thermal energy
to preheat food for PEF treatment at moderate (inlet) temperature (e.g., 40°C). A combination of
mild heat and PEF might also be helpful to achieve sufficient enzyme inactivation to avoid the
necessity of refrigerated storage. When operating at high treatment temperatures and making use
of synergetic heat effects, PEF energy input might be reduced close to the amount required for
conventional thermal pasteurization, assuming 95% of heat recovery (Toepfl et al., 2006). Despite
of increase in delivered electrical power, PEF pasteurization seems to be less energy-intensive than
traditional pasteurization methods, it leads to annual savings of 791.2–1055 TJ per year of fossil
fuel-equivalents, which also contribute in reduction of CO2 emissions (Lelieveld, 2004).
The study (Lung et al., 2006) provided estimated information on the potential energy
savings of PEF and dielectric drying systems compared to existing technologies. The electricity
savings of PEF can be up to 18%, based on the assumed electricity consumption range of the
base technology. Similarly, the combined application of high pressure and heat can be utilized
to achieve an inactivation of spores of Clostridium spp. like that of conventional sterilization.
However, the specific energy input required for sterilization of cans can be reduced from 300 to
270 kJ/kg when applying the HHP treatment. In case of HHP processing, a compression energy
recovery rate of 50% can be estimated when a two-vessel system or pressure storage is used,
by making use of energy recovery, a specific energy input of 242 kJ/kg will be required for
sterilization, corresponding to a reduction of 20% in the total energy requirements (Toepfl et al.,
2006).
Allowing a precise control of the process temperature leads to lower energy costs, reduced
gas consumption and less combustion-related emissions in PEF (Wang, 2014; Lima et al., 2002;
Nowak and Lewicki, 2004). Also, the pressure-assisted sterilization processes in HHP application
is considered to be a waste-free process. In fact, the pre-sterilization of packaging, e.g., H2O2 or
other chemical agents will not be required and will therefore contribute to a reduction of the
amount of chemicals in the liquid effluents.
In order to achieve their full potential for industrial implementation and commercial exploi-
tation, issues related with the environmental impact, such as, wastewater and gas emissions, the
conservation of non-renewable resources and energy consumption are increasingly attracting the
attention of the food processors since they can represent significant reductions of the processing
costs.
The novel non-thermal technologies for food processing provide not only energy savings, but
also water savings, increased reliability, reduced emissions, higher product quality, improved
productivity (Masanet et al., 2011), and consequently, less impact on the environment. In fact, it is
estimated that in the food industry ca. 57% of the fossil fuel consumption is used to generate steam
(Einstein et al., 2001).
It seems clear that emerging thermal and non-thermal technologies have clear environmental
benefits, by improving the overall energy efficiency of the process or by reducing the use of
non-renewable resources. A review on the environmental aspects of nonthermal preservation
technologies concluded that these technologies do not generate contaminants, and thus have the
potential to be considered clean technologies (Pereira and Vicente, 2010). Most of the food industry
uses indirect energy, such as steam raising account for the biggest share of the combustion process
emissions. The combustion processes release a range of pollutants (smoke, carbon dioxide and
organic compounds). These emissions largely dependent on the type of fuel used (Toogood and
Key, 2000). Therefore, food processing organizations powered only by electricity might present an
environmental advantage when compared with traditional processing technologies. In general, the
novel technologies are considered environmentally friendly. They may eliminate completely, or at
least reduce significantly, the local use of boilers or steam generation systems and consequently,
diminish waste water and increase water and energy savings.
20.4 paCKaGING ISSUES
Non-thermal processes can be used in aseptic processing systems to replace heating units. But
utmost care should be taken to package the food properly and avoid loss of sterility. Packaging mate-
rials that are compatible with the non-thermal processing and do not undergo adverse changes affect-
ing the quality of the processed product should be selected (Brody, 2005). The initial high quality of
non-thermally processed food can be maintained during the storage period to a great extent if proper
packaging materials are selected. The packaging materials used for non-thermally processed food
will have different properties and requirements when compared to the ones used for food subjected
to conventional heat treatment (Han, 2008). One of the greatest advantages of low-temperature pro-
cessing is that there is no requirement for packaging materials with high melting temperatures. Cold
sealing of packages can be employed using various adhesives, polymers and sealants that produce
less volatiles and require less printing solvents (Morris et al., 2007). Commercialization of non-
thermal processing of food can be made possible only if there is proper understanding of packaging
requirements of the product, characteristics of the packaging material and consumer response and
acceptance of the packaged non-thermally processed food. Firstly, the special requirements of pack-
aging materials for non-thermal processing should be identified. For this, process parameters and
microbial death kinetics should be studied in detail (Balasubramaniam et al., 2004; Brody, 2007).
The packaging requirements will vary depending on the non-thermal method used. In the case
of high-pressure processing, the packaging material used should be flexible and should be able to
withstand the compression forces without losing their physical integrity (Balasubramaniam et al.,
2004). Glass and metal containers and paperboards are not suitable HPP foods (Caner et al., 2004).
Packaging materials composed of ethylene vinyl alcohol (EVOH) and polyvinyl alcohol (PVOH)
are found to be compatible with high-pressure treatment (Masuda et al., 1992). Oxygen permeability
of packaging materials is a very important factor of oxidation of the foods. HPP should use high-
oxygen-barrier packaging materials. Also, it is important to keep the headspace small in order to
avoid deformation strain on the packaging material as air is compressible under pressure (Lambert
et al., 2000). A common problem encountered is the seal defect or delamination of the package post
high-pressure processing which needs to be addressed. Special designs for securing the seals and
preventing delamination should be established.
Irradiation is commonly used to sterilize packages in aseptic processing. In case of cold

sterilization of food by irradiation, packaging is done prior to the treatment in order to avoid
recontamination. There are a few concerns about the effect of irradiation on the packaging
material characteristics, which are found to alter post-treatment. Permeability of gases change
after irradiation and this can have detrimental effect on the quality of the product during storage.
Radiolysis products of irradiation such as peroxides and free radicals from food as well as volatile
hydrocarbons produced from polymers or additives can degrade the quality of both packaging
material and food (Barbosa-Canovas et al., 1998; Crook and Boylston, 2004). Hence, any material
used should be subjected to toxicological tests after irradiation and only then used as a commercial
packaging material.
For PL-processed products the packaging material should be transparent and UV transmissive.
Generally most food packaging will have good UV barrier properties. A practical difficulty in
designing packaging for PL-treated product is that printing on packages can cause non-uniform
exposure of product surface to the treatment as decontamination by PL is done after packaging.
Moreover, glass and PET containers cannot be used for packing products as they absorb UV
radiations (Brody, 2008).
Aseptic food packaging is found to be most suitable for PEF-processed food. Some PEF-
processed foods show quality problems during storage due to higher rate of oxidative reactions when
compared to thermally processed foods. Therefore, packaging materials considered should have very
high-oxygen barrier property. The quality of PEF-processed food, especially juices, is found to be
very high. By selecting proper packaging material, such high initial quality can be extended and
maintained over time. There is a need for more studies and investigations to understand the oxidation
kinetics in PEF-treated food and accordingly design suitable packaging systems (Morris et al., 2007).
Consumer acceptance of non-thermally processed food plays a critical role in commercializa-
tion of the technology. In this regard, packaging can significantly influence the consumers reaction
to new processing technology (Han, 2008). For the non-thermal processes that consumers are not
familiar with, food packaging materials should provide information on the new processes to reduce
the reluctance among consumers. Sticking to the old design and structure of the package, without
much alteration, when using a new technology could make the consumers less reluctant to buy the
product. New and unfamiliar package design for a non-conventionally processed food can lead to
scepticism (Raso and Barbosa-Cánovas, 2003; Brody, 2005).
To facilitate commercialization of non-thermally processed food, various regulations should be
established. Industries and regulatory agencies should collaborate with scientists to develop testing
and risk assessment protocols for the packaging materials so that they can be safely implemented
for non-thermally processed food (Morris et al., 2007).
20.5 CONSUMEr rESpONSE
Consumers today, are well aware of the detrimental effects of thermal processing on food qual-
ity. One of the major factors that has spurred interest for non-thermal food processing and driven
industries to accept and implement these alternate techniques is increasing consumer demand for
food products which are free of chemicals and are “minimally” or gently processed (Knorr et al.,
2002). The commercial success of any food processing technology is highly depended on the con-
sumers’ acceptance. Good sensory and nutritional profiles, although highly desired, will not guar-
antee success in the market. This is because consumers today make their food choices not only
based on the product attributes but they are also influenced by factors such as origin, price, animal
welfare and technology used. Genetically modified and irradiated foods are best examples where
the advantages of the technology did not render commercial success mainly due to public’s aversion
and fear towards them (Nielson et al., 2009).
Irradiation, although, emerged as a non-thermal technology almost a century ago, still struggles
to gain public acceptance, despite its numerous benefits and effectiveness in increasing shelf life of
many food. A number of studies were conducted on the public acceptance of this technology in the
1980s and 1990s. Majority of these studies found that the negative response for irradiation breeds
from unfamiliarity and lack of knowledge about the technology (Frewer et al., 2011). The general
public is concerned about the formation of perceived “carcinogenic” compounds in irradiated food
and health hazards to workers and those handling the equipment. Such a response has critically
hampered the widespread implementation of irradiation by food industries. Interestingly, recent
studies have noted that the use of the term “ionizing energy” instead of “irradiation” invoked much
less concern among consumers, suggesting the name of the technology also plays a critical role in
its acceptance by the public (Cardello, 2003).
High-pressure-processed products have gained commercial marketability as consumers per-
ceived the products naturalness, high nutrient content and improved taste and texture as highly
advantageous. They considered the environmental friendliness of the technology while evaluating
HPP- and PEF-processed products and saw it as a major advantage. However, lack of information
about the technology and higher price of the products were causes for concern. In case of PEF-
processed products, there was low awareness and initial scepticism, as consumers associated this
method with using electricity in food and were concerned of its unknown side effects (Nielson et al.,
2009; Frewer et al., 2011).
Thus, creating public awareness about the non-thermal technologies and providing them
information through widespread campaigns can help in gaining consumer acceptance for products
prepared using these technologies. It is important to note that, regardless of the safety and quality
of the product, consumer familiarity with a technology governs its commercialization, as food is a
critical consumer product is any culture or society (Han, 2008).
20.6 rEGULatOrY aSpECtS
Official permission from governmental regulatory agencies to use the new technology is one of
the requirements for the commercialization of new processes. Since food-related legislation process
is very political, the regulatory issues of the permission to use any non-thermal process are also
political. Food industry, consumer groups, nonthermal equipment manufacturing industries and
regulatory agencies may have different voices to the same issue. Furthermore, there are not enough
scientific data and results to provide one solution to these diverged parties of interests. Therefore, it
is very important to identify the current development status of technology commercialization and
for scientific societies and academic institutes to provide a fair milestone to these interested groups.
Although PL is not widely used in food industry, it has been successfully used in large-
scale commercial application for CVD/CD/Blu-Ray bonding and bottle cap decontamination.
Commercial-scale food packaging decontamination system based on PL was commissioned by
FDA in 1995. Therefore, these advanced, large-scale commercial systems can be easily adapted
to food industry. PL technology can also easily be integrated into existing food processing lines,
making it an ideal technology.
PEF is a physical process used to preserve food, and thus it falls under the US Food and Drug
and Administration (FDA) regulations. The difficulty experienced in demonstrating the nonthermal
methods as good as thermal processes is slow commercialization of new preservation technologies.
Before the commercialization of processing of foods by nonthermal technology must comply with
appropriate safety regulations set forth by the FDA according to the type of product. FDA regula-
tions have been established to assure the mission of protecting public health. However, the FDA
does not approve processes per se, rather the use of substance or components used in the process.
Besides complying with current regulations that might apply, PEF processing may also require
premarket approval. This is due to the use of food contact surfaces under different conditions (pres-
ervation techniques). In PEF treatment chamber, electrode material could possibly migrate to the
food. To determine if these of other aspects are safety concerns for a PEF process, a petition must
be submitted to FDA for evaluation. After the evaluation, a ruling will be made and issued and
then included in the codex of federal regulations (CFR). In the premarket approval process, PEF
processors are asked to submit a scientific review with necessary data to clearly characterize the
process and the product. The information package submitted to the FDA should address the fol-
lowing: a) type of product or process being evaluated, b) how the product will be used, and c) what
the product will accomplish. The environmental impact of the product is also important (Hansen,
1999). However, similar dialog can be established to demonstrate that a petition is not needed for
premarket approval, as was the case in 1997, when pure pulse received a letter of no objection, and
therefore no rulemaking was required. Each processor must follow the same procedure through
intense communication with the FDA. When a new process is filed, it is necessary to a) established
an active and continuous dialog with the FDA during process developed (Larkin and Spinak, 1996),
b) meet with the FDA to describe the process, c) invite the FDA for an on-site visit (pilot and produc-
tion facility), d) draft and provide the FDA with an outline of the proposed filing, and e) identify the
most resistant organism to commercial viability and the least lethal treatment zone in the system.
Regarding the novelty of the process, the FDA will be interested in reviewing equipment design.
Four aspects should address concerns in area of toxicology, nutrition, and microbiology.
Information submitted must address the following questions: a) does the process generate chemical
changes producing compounds that may be toxic, b) are there nutrient losses, and c) if the final
product is not sterile, how will the product or process control certain microorganism and production
of toxin. Today, the basis for implementing this technology exists and is well supported by several
decades of research. Now is the time for systematic research that addresses specific FDA technology
exists and is well supported even after several decades of research. Now is the time for systematic
research that addresses specific FDA questions and other issues regarding implementation of PEF
in production line.
FDA regulatory control may be specific to the US but the relevance of delivering safe foods to
consumers goes beyond US borders. In US, the FDA has approved the use of ozone like antimicrobial
agent for the direct contact with all foods and food products (June 26, 2001). The Ministry of
Agriculture of the United States accepted ozone similar to antimicrobial agent for the direct contact
with meats, poultry, fish, molluscs, and crustaceans (December 2001). The government of Japan
in 1996 admitted its use for the direct contact with all types of food. Ozone is listed by FDA as
“generally recognized as safe” (GRAS). The Japanese fishing boats use ozonized water routinely to
wash the fresh fish and to make ice with ozonized water and pack it on board. The Canadian Food
Inspection Agency (CFIA) has approved the use of ozone for the cleaning the contact surfaces of
food. The government of Australia (in 1996) authorized the use of ozone for the contact with all
foods—similar to the Japanese approval. The German fishing boats also use ozone in the water
and in ice. In Norway, the farms of aquaculture and the plants of processing use ozone for the
preservation of the fish.
More than 40 countries have approved 50 and above food for commercial irradiation for
various purposes and marketing. Established by national health authorities each country has its
own regulations. For example, food irradiation was considered as food additive by US Congress in
Merrill (1997). Some countries established regulation based on a case-by-case basis. Many other
countries used FAO/IAEA/WHO Expert Committee on the Wholesomeness of Irradiated Food
(JECFI) provided facts, conclusions, and recommendations as their references. However, the food
items and purpose of irradiation vary from country to country. The concept of “chemiclearance”
accepted by the U.S. FDA to approve irradiation of fruits or meat was a milestone in rule-making
for irradiated foods. In the U.S., all irradiated foods (except spices) required to be labelled, with
a “radura” logo, and phrasing such as “treated with radiation to control spoilage” or “treated by
irradiation for quarantine purpose.” Manufacturers producing irradiated foods can identify the
radiation source used, such as “treated with gamma-radiation,” “treated with ionizing radiation,”
or “treated with x-radiation.” They can additionally add statements as part of a consumer education
effort such as “this treatment does not induce radioactivity” if the statements are truthful and not
misleading to the consumers.
Microbiological criteria for safe production of foods by HPP needs to be established
(Balasubramaniam et al., 2002). In the United States, the Food, Drug, and Cosmetic Act (FD&C
Act), which requires all foods to be processed, packaged, and held under sanitary conditions, is the
basis by which FDA promulgates specific regulations. Currently, high-pressure pasteurized product
(oysters), distributed under refrigerated conditions, similar to thermally pasteurized products. high-
pressure pasteurized products are mandatory to be processed under GMP conditions and relevant
commodity specific regulations (e.g., juice HACCP, Pasteurized Milk Ordinance (PMO), Sea Food
HACCP etc.). The potential for temperature exploitation during refrigerated storage and distribution
must be carefully evaluated and minimized. Processors must also work with equipment vendors to
ensure that any part of the pressure vessel, which might have incidental contact with the food, is only
made from approved materials. Currently, high-pressure sterilized low-acid shelf stable products
are not commercially sold in the United States. In European countries, the national regulations
related to the application of the precautionary principle for new products have been replaced by
an EU regulation for novel foods and ingredients (CE258/97), which came into force in 1997. This
legislation for “novel foods” establishes an evaluation and a license system, compulsory for all new
foods and processes. To simplify the regulation, it was recently admitted that any new product could
be treated at a national regulation level, if it is possible to show that the product is substantially
equivalent to a product already on the market (Rastogi et al., 2007).
The regulation is the most critical factor for the use of nonthermal processes as it can either
facilitate or limit the use of non-thermal processes. Guidelines on the operation conditions of non-
thermal processes, compatibility of selected packaging materials with the technology, and more
evidence on the safety, functionality, and nutritional quality are needed to convince regulatory
authorities that these non-thermal-processed products are safe to be commercially made available
to the public (Han, 2008).
20.7 CUrrENt StatUS OF COMMErCIaL appLICatIONS
Over the years, as favourable laboratory research findings on the effect of non-thermal process-
ing on food quality, shelf life, packaging, and pathways of microbial spoilage emerged, there was a
need to facilitate technology transfer to the food processing industry and commercialize these tech-
nologies. The major drawback that slowed the commercialization of many non-thermal processing
technologies was the lack of systems and equipments for large-scale application. Equipments that can
operate during multiple shifts at low cost, are easy to maintain and repair and have high productivity
to yield economical returns to the food processor were needed to be manufactured. Also, pilot-scale
food processing lines to prepare samples for storage studies and consumer testing were needed.
Extensive research in high-pressure processing of food around the world provided the basic
science needed for the commercial implementation of this the technology. Guacamole made from
avocados by the US food company Avomex (presently Fresherized Foods) and jams, jellies, and
sauces produced by Meidi-ya Food Co in Japan were among the first high-pressure-processed
product marketed to consumers in the early 1990s. Several high-pressure vessel manufacturers like
Uhde in Germany; ABB Pressure Systems (Quintus) in Sweden; Engineered Pressure Systems, a
division of National Forge, in Belgium and the United States; ACB GEC, in France; and Stansted
Fluid Power, in the United Kingdom showed interest in developing and promoting high-pressure
processing of food (Asaka and Hayashi, 1991; Ledward et al., 1995).
High-pressure processing has found applicability in the processing of a variety of product cate-
gories including cooked meats, seafood and fish, vegetables, and fruit juice. In the last two decades,
large HPP research programs have been established in Japan, Europe, and the United States of
America (Torres and Velazquez, 2005). It has now become a commercially implemented technol-
ogy with around 82 commercial-scale high-pressure food processing systems in use worldwide with
an estimated production of 150,000 tons/year and sales over $2 billion annually in the United States
alone (Pereira and Vicente, 2010; Bermúdez-Aguirre and Barbosa-Cánovas, 2011). The list of coun-
tries processing and selling high-pressure products is also growing fast; these include Canada, USA,
Mexico, Colombia, Chile, Brazil, Ireland, United Kingdom, Norway, Finland, Poland, Germany,
France, Italy, Spain, Portugal, India, Korea, Japan, Australia, and New Zealand (Bermúdez-Aguirre
and Barbosa-Cánovas, 2011).
As far as the cost of pressurized products is concerned, the current availability of high-pressure
equipment has reduced the cost of products processed by this technology significantly in recent
years, making more products accessible to consumers. According to Rastogi et al. (2007), the aver-
age cost of high-pressure processing is around US$0.05–0.5/L or kg depending on processing con-
ditions, which is lower than thermal processing costs. However, according to Hernando Saiz et al.
(2008) the cost of high-pressure processing under some common processing conditions, for exam-
ple, ready-to-eat meats (585 MPa, 3 min, filling vessel 50%), is really between US$0.08 and 0.22/kg,
which makes HHP technology suitable for products of premium quality. Some of the technological
barriers in developing HPP vessel are the need for wire winding (for safe and reliable operation) that
increases the equipment cost and unavailability of pressure vessels that can operate above 680 MPa.
HPP products rely on refrigeration, reduced water activity or low pH to prevent bacterial spore out-
growth (Ting et al., 2002; Torres and Velazquez, 2005).
PEF, however, has not been able to find similar commercial success, in spite of all research
and development. PEF implementations mostly lie at the experimental level and in the pilot plant
range. Major suppliers of commercial-scale equipment are Elea (Quakenbruck, Germany) and
Pulsemaster (Ladel, the Netherlands), specializing in PEF processing for mild preservation. The
first commercial PEF application was implemented in 2005 by Genesis Juice Corporation (USA)
in 2005 implemented PEF for the first time for nonthermal pasteurization of fruit juice in a 200L/h
processing unit (Clark, 2006). PEF was found to reduce loss of flavour and improve the shelf life
of the product. The current design of PEF treatment chambers does not allow processing of solid
foods and thus limits its use for a wide range of products. High capital cost of the equipment
is another barrier that has hampered the commercialization of this technology. In the current
state of the technology, electrodes need replacement after about 100 h of operation. Chamber
design related to efficiency is still affecting large-scale operation. As fluid volumes increase, larger
diameter tubing will be required inside the chambers. Lager tubing implies larger gaps in the
treatment chambers resulting in higher voltages and energy consumption. Improving electrode
reliability and optimizing chamber design are issues that need to be tackled in order to promote
the widespread use of PEF as a pasteurization technique (Ortega-Rivas, 2011). On a brighter note,
for liquid food, PEF is a rapid treatment that is well adapted to continuous processing. Maximal
PEF treatment intensity is limited by equipment design and food’s ability to withstand dielectric
breakdown (Lado and Yousef, 2002).
Irradiation of drugs and medical devices was practiced by pharmaceutical, plastics, and
packaging industries starting in the 1950s. However, public aversion towards this technology mainly
on the grounds of safety and possible formation of carcinogenic compounds in treated food had
retarded the commercial application of this technology for food sterilization for almost a century
since its emergence (Baily, 1957). In recent times, with increased public awareness, irradiation of
fruits and vegetables to prevent the spread of pest and diseases has been increasing globally. In
2010, 18,446 tonnes of fruits and vegetables were irradiated in Mexico, US, Thailand, Vietnam,
Australia, and India for export (Sehrawat et al., 2018).
PL has been in commercial use for disinfecting food contact surfaces for quite some time
now. PL equipment that are being manufactured today have capacity to sterilize food packages
at a rate of 40,000 units/h (Riedel, 2007). Major manufacturers and suppliers of PL equipment
include Claranor (France). Commercial application of ultrasound was hindered by several factors
such as cost of the equipment and energy requirements until recent times. However, manufacturers
of ultrasound equipment have been focusing on developing systems to reduce operating cost by
designing multiple systems in series and parallel that facilitate larger flow rates (Yuting et al.,
2013). Current equipments have energy required per litre material treated comparable to any other
unit operation in the industry, are robust and durable with only probes which are in direct contact
requiring replacement every 18 months (Patist and Bates, 2008).
Ozone and UV radiation have been used worldwide for supplying drinking water. More than
500 UV plants for disinfecting drinking water operate in North America, and in Europe more than
2000 plants use this technology (Pereira and Vicente, 2010). The capacity of ozone generators varies
depending on the application. Large units treat more than 100,000 L/h in municipal water supplies.
Small, countertop units treat 1,500 L/h in labs and food processing plants (e.g., cutting boards,
utensils, and hand sanitizing).
20.8 CONCLUSION
Non-thermal food processing technologies have been the focus of numerous research studies and
investigations in past couple of decades. Their impact on food chain security is immense and they
have huge potentials to improve the overall quality of processed foods. Application of these emerg-
ing technologies for recovery of valuable components from food wastes is another promising field
which requires due attention. While certain factors have limited the commercialization of some non-
thermally processed food, some other non-thermal technologies have received favourable response
in the market. Cost-effectiveness is a major concern that needs to be addressed. Some NTP tech-
nologies are relatively more costly than others. Most of the technologies are found to be environ-
ment friendly and facilitate energy savings when compared to thermal food processing. One of the
biggest barriers to commercialization of non-thermal technologies is undoubtedly negative response
from consumers. Technologies like irradiation and pulsed electric field are not well perceived, lead-
ing to rejection of treated products by consumers. Most of the time, such a response breeds from
misunderstandings and unfamiliarity with the working of the technology. Because of lack of proper
knowledge, they relate these technologies to harmful events and toxicity and assume that the non-
thermally processed products will be expensive. Therefore, consumer education to increase aware-
ness on the applications of non-thermal technologies for food preservation is of utmost importance
and crucial to the successful commercialization of treated food products. Besides consumer, accep-
tance has to also come from food processors, equipment manufacturers and regulatory bodies. The
use of nonthermal processes for food preservation will become more popular and common in the
future. However, the concerns regarding the various scientific and non-scientific parameters should
be addressed and resolved through communication and collaboration between industry and aca-
demia and thus accelerate the rate commercialization of non-thermally processed food.
rEFErENCES
Alighourchi, H., Barzegar, M., and Abbasi, S. (2008). Effect of gamma irradiation on the stability of antho-
cyanins and shelf-life of various pomegranate juices. Food Chem. 110: 1036–1040.
Alothman, M., Bhat, R., and Karim, A. A. (2009a). Effects of radiation processing on phytochemicals and
antioxidants in plant produce. Trends Food Sci Technol. 20: 201–212.
Alothman, M., Bhat, R., and Karim, A. A. (2009b). UV radiation-induced changes of antioxidant capacity of
fresh-cut tropical fruits. Innov Food Sci Emerg Technol.10: 512–516.
Asaka, M., and Hayashi, R. (1991). Activation of polyphenoloxidase in pear fruits by high pressure treatment.
Agric Biol Chem. 55: 2439–2440.
Baily, N. A. (1957). Patient exposure to ionizing radiation in dental radiography. Radiology. 69: 42–45.
Balasubramaniam, V. M., Sizer, C. E., and Ting, E. (2002). Validating high-pressure processes for low-acid
foods. Food Technol. 56: 36–42.
Balasubramaniam, V. M., Ting, E. Y., Stewart, C. M., and Robbins, J. A. (2004). Recommended labora-
tory practices for conducting high-pressure microbial inactivation experiments. Innov Food Sci Emerg
Technol. 5: 299–306.
Barboni, T., Cannac, M., and Chiaramonti, N. (2010). Effect of cold storage and ozone treatment on physi-
cochemical parameters, soluble sugars and organic acids in Actinidia deliciosa. Food Chem. 121(4):
946–951.
Barbosa-Cánovas, G. V., Gongora-Nieto, M. M., and Swanson, B. G. (1998). Nonthermal electrical methods in
food preservation/Metodos electricos no termicos para la conservacion dealimentos. Food Sci Technol
Int. 4: 363–370.
Bermúdez-Aguirre, D., and Barbosa-Cánovas, G. V. (2011). An update on high hydrostatic pressure, from the
laboratory to industrial applications. Food Eng. 3: 44–61.
Bhat, R., and Sridhar, K. R. (2008). Nutritional quality evaluation of electron beam-irradiated lotus (Nelumbo
nucifera) seeds. Food Chem. 107: 174–184.
Bhat, R., Sridhar, K. R., and Tomita-Yokotani, K. (2007). Effect of ionizing radiation on antinutritional fea-
tures of velvet bean seeds (Mucuna pruriens). Food Chem. 103: 860–866.
Brody, A. L. (2005). Packaging for nonthermally and minimally processed foods. Food Technol. 59(10): 75–77.
Brody, A. L. (2007). The role of active packaging in nonthermal processing systems. In: Packaging for
Nonthermal Processing of Food. pp. 17–32. Han, J.H., Ed., Blackwell Publishing, Ames, IA.
Butz, P., and Tauscher, B. (2002). Emerging technologies: Chemical aspects. Food Res Int. 35: 279–284.
Caner, C., Hernandez, R. J., and Harte, B. R. (2004). High‐pressure processing effects on the mechanical,
barrier and mass transfer properties of food packaging flexible structures: A critical review. Packag
Technol Sci. 17: 23–29.
Cardello, A. V. (2003). Consumer concerns and expectations about novel food processing technologies: Effects
on product liking. Appetite. 40: 217–233.
Cheng, L. H., Soh, C. Y., Liew, S. C., and Teh, F. F. (2007). Effects of sonication and carbonation on guava
juice quality. Food Chem. 104: 1396–1401.
Chipurura, B., and Muchuweti, M. (2010). Effect of irradiation and high pressure processing technologies on
the bioactive compounds and antioxidant capacities of vegetables. Asia Pac J Clin Nutr. 2: 190–199.
Clark, J. P. (2006). Processing-pulsed electric field processing. Food Tech. 60(1): 66–67.
Crook, L. R., and Boylston, T. D. (2004). Flavor characteristics of irradiated apple cider during storage: Effect
of packaging materials and sorbate addition. J Food Sci. 69: 557–563.
Cullen, P. J., Valdramidis, V. P., Tiwari, B. K., Patil, S., Bourke, P., and O’donnell, C. P. (2010). Ozone process-
ing for food preservation: An overview on fruit juice treatments. Ozone: Sci Eng. 32: 166–179.
Einstein, D., Worrell, E., and Khrushch, M. (2001). Steam systems in industry: Energy use and energy
efficiency improvement potentials. No. LBNL-49081. Ernest Orlando Lawrence Berkeley National
Laboratory, Berkeley, CA.
Frewer, L. J., Bergmann, K., Brennan, M., Lion, R., Meertens, R., Rowe, G., and Vereijken, C. M. J. L. (2011).
Consumer response to novel agri-food technologies: Implications for predicting consumer acceptance
of emerging food technologies. Trends Food Sci Technol. 22: 442–456.
Guerrero-Beltran, J. A., and Barbosa‐Canovas, G. V. (2005). Reduction of Saccharomyces cerevisiae,
Escherichia coli and Listeria innocua in apple juice by ultraviolet light. J Food Process Eng. 28:
437–452.
Han, J. J. H. (2009). Food safety and innovative food packaging. In: Microbiologically Safe Foods, pp.
507–521, Heredia, N., Wesley, I., and Garcia, S., Eds., John Wiley & Sons, New York.
Hansen, L. G. (1999). Environmental regulation through voluntary agreements. In: Voluntary Approaches in
Environmental Policy, pp. 27–54. Carraro, C., and Lévêque, F., Eds., Springer, Dordrecht.
Hendrickson, J. (1996). Energy Use in the US Food System: A Summary of Existing Research and Analysis.
Madison, WI: Center for Integrated Agricultural Systems.
Hernando Saiz, A., Tarrago Mingo, S., Purroy Balda, F., and Tonello Samson, C. (2008). Advances in design
for successful commercial high pressure food processing. Food Australia. 60: 154–156.
Karaca, H., and Velioglu, Y. S. (2007). Ozone applications in fruit and vegetable processing. Food Rev Int.
23: 91–106.
Knorr, D., Ade-Omowaye, B. I. O., and Heinz, V. (2002). Nutritional improvement of plant foods by non-
thermal processing. Proc Nutr Soc. 61: 311–318.
Lado, B. H., and Yousef, A. E. (2002). Alternative food-preservation technologies: Efficacy and mechanisms.
Microbes Infect. 4: 433–440.
Lambert, Y., Demazeau, G., Largeteau, A., Bouvier, J., Laborde-Croubit, S., and Cabannes, M. (2000).
Packaging for high-pressure treatments in the food industry. Packag Technol Sci. 13: 63–71.
Larkin, J. W., and Spinak, S. H. (1996). Safety considerations for ohmically heated, aseptically processed,
multiphase low-acid food products. Food Technol. 50(5): 242–245.
Ledward, D. A. (1995). High Pressure Processing of Foods. Loughborough: Nottingham University Press.
Lee, S. K., and Kader, A. A. (2000). Preharvest and postharvest factors influencing vitamin C content of hor-
ticultural crops. Postharvest Biol Technol. 20: 207–220.
Lelieveld, H. (2004). PEF—A food industry’s view. In Novel Food Processing Technologies, pp. 190–202.
Barbosa-Cánovas, G. V., Tapia, M. S., and Cano, M. P., Eds., CRC Press, Boca Raton, FL.
Lima, M., Zhong, T., and Lakkakula, N. R. (2002). Ohmic heating: A value-added food processing tool. La
Agr. 45: 16–16.
Lung, R. B., Masanet, E., and McKane, A. (2006). The role of emerging technologies in improving energy
efficiency: Examples from the food processing industry. In Proceedings of the Industrial Energy
Technologies Conference, New Orleans, LA.
Masanet, E. R., Brown, R. E., Shehabi, A., Koomey, J. G., and Nordman, B. (2011). Estimating the energy use
and efficiency potential of US data centers. Proceedings of the IEEE. 99: 1440–1453.
Masuda, M., Saito, Y., Iwanami, T., and Hirai, Y. (1992). Effects of hydrostatic pressure on packaging mate-
rials for food. Colloques-Institut National De La Sante Et De La Recherche Medicale Colloques Et
Seminaires, pp. 545–545.
Merrill, R. A. (1997). Food safety regulation: Reforming the Delaney Clause. Annu Rev Public Health 18(1):
313–340.
Morales-de La Peña, M., Salvia-Trujillo, L., Rojas-Graü, M. A., and Martín-Belloso, O. (2010a). Impact of
high intensity pulsed electric field on antioxidant properties and quality parameters of a fruit juice–
soymilk beverage in chilled storage. LWT-Food Sci Technol. 43: 872–881.
Morales-De La Peña, M., Salvia-Trujillo, L., Rojas-Graü, M. A., and Martín-Belloso, O. (2010b). Isoflavone
profile of a high intensity pulsed electric field or thermally treated fruit juice-soymilk beverage stored
under refrigeration. Innov Food Sci Emerg Technol. 11: 604–610.
Morris, C., Brody, A. L., and Wicker, L. (2007). Non‐thermal food processing/preservation technologies: A
review with packaging implications. Packag Technol Sci. 20: 275–286.
Nielsen, H. B., Sonne, A. M., Grunert, K. G., Banati, D., Pollák-Tóth, A., Lakner, Z., and Peterman, M. (2009).
Consumer perception of the use of high-pressure processing and pulsed electric field technologies in
food production. Appetite. 52: 115–126.
Nowak, D., and Lewicki, P. P. (2004). Infrared drying of apple slices. Innov Food Sci Emerg Technol. 5:
353–360.
Ortega-Rivas, E. (2011). Critical issues pertaining to application of pulsed electric fields in microbial control
and quality of processed fruit juices. Food Bioprocess Technol. 4: 631–645.
Patist, A., and Bates, D. (2008). Ultrasonic innovations in the food industry: From the laboratory to commer-
cial production. Innov Food Sci Emerg Technol. 9: 147–154.
Pereira, R. N., and Vicente, A. A. (2010). Environmental impact of novel thermal and non-thermal technolo-
gies in food processing. Food Res Int. 43: 1936–1943.
Prasad, K. N., Yang, E., Yi, C., Zhao, M., and Jiang, Y. (2009). Effects of high pressure extraction on the
extraction yield, total phenolic content and antioxidant activity of longan fruit pericarp. Innov Food Sci
Raso, J., and Barbosa-Cánovas, G. V. (2003). Nonthermal preservation of foods using combined processing
techniques. Cr Rev Food Sci. 43: 265–285.
Rastogi, N. K., Raghavarao, K. S. M. S., Balasubramaniam, V. M., Niranjan, K., and Knorr, D. (2007).
Opportunities and challenges in high pressure processing of foods. Crit Rev Food Sci Nutr. 47: 69–112.
Riedel, C. (2007). Pulsed light treatment for food and packaging. Presentation at novel Q seminar, 9–10 July,
held at Campden BRI, Chipping Campden, Gloucestershiree.
Rodriguez‐Gonzalez, O., Buckow, R., Koutchma, T., and Balasubramaniam, V. M. (2015). Energy require-
ments for alternative food processing technologies—Principles, assumptions, and evaluation of effi-
ciency. Compr Rev Food Sci Food Saf. 14: 536–554.
Sajilata, M. G., and Singhal, R. S. (2006). Effect of irradiation and storage on the antioxidative activity of
cashew nuts. Radiat Phys Chem. 75: 297–300.
Sanchez-Moreno, C., De Ancos, B., Plaza, L., Elez-Martinez, P., and Cano, M. P. (2009). Nutritional approaches
and health-related properties of plant foods processed by high pressure and pulsed electric fields. Crit
Rev Food Sci Nutr. 49: 552–576.
Sehrawat, R., Khan, K. A., Goyal, M. R., and Paul, P. K. (Eds.). (2018). Technological Interventions in the
Processing of Fruits and Vegetables. CRC Press.
Stute, R., Klingler, R. W., Boguslawski, S., Eshtiaghi, M. N., and Knorr, D. (1996). Effects of high pressures
treatment on starches. Starch‐Stärke. 48: 399–408.
Ting, C. E., Balasubramaniam, E., and Rasanayagam, V. (2002). Determining thermal effects in high-pressure
processing. Food Tech. 56: 31–35.
Toepfl, S., Mathys, A., Heinz, V., and Knorr, D. (2006). Potential of high hydrostatic pressure and pulsed elec-
tric fields for energy efficient and environmentally friendly food processing. Food Rev Int. 22: 405–423.
Toogood, S., and Key, M. (2000). Air pollution. In: Food Industry and the Environment in the European
Union, Practical Issues and Cost Implications. Dalzell, J. M., Ed., Gaithersburg, MD: Aspen Publishers
Inc.
Torres, J. A., and Velazquez, G. (2005). Commercial opportunities and research challenges in the high pres-
sure processing of foods. J Food Eng. 67: 95–112.
Wang, L. (2014). Energy efficiency technologies for sustainable food processing. Energy Efficiency. 7: 791–810.
Whangchai, K., Saengnil, K., and Uthaibutra, J. (2006). Effect of ozone in combination with some organic
acids on the control of postharvest decay and pericarp browning of longan fruit. Crop Prot. 25: 821–825.
Wong, P. Y., and Kitts, D. D. (2001). Factors influencing ultraviolet and electron beam irradiation-induced free
radical damage of ascorbic acid. Food Chem. 74: 75–84.
Wood, O. B., and Bruhn, C. M. (2000). Position of the American Dietetic Association: Food irradiation.
J Acad Nutr Diet. 100: 246.
Yen, G. C., and Lin, H. T. (1996). Comparison of high pressure treatment and thermal pasteurization effects
on the quality and shelf life of guava puree. Int J Food Sci Technol. 31: 205–213.
Yuting, X., Lifen, Z., Jianjun, Z., Jie, S., Xingqian, Y., and Donghong, L. (2013). Power ultrasound for the
preservation of postharvest fruits and vegetables. Int J Agric Biol Eng. 6: 116–125.
ChaptEr 21
Consumer acceptance and Future trends

of Non-thermal-processed Foods
Prerna Nath, S. J. Kale, and Bharat Bhushan
CONtENtS
21.1 Introduction ........................................................................................................................ 433

21.2 Food Irradiation.................................................................................................................. 434
21.3 High-Pressure Processing .................................................................................................. 437
21.4 Pulsed Electric Field .......................................................................................................... 443
21.5 Cold Plasma Technology ....................................................................................................444
21.6 Consumer Responses to Recent Innovations in Food Processing Technologies................ 445
21.7 Conclusions ........................................................................................................................ 450
References ...................................................................................................................................... 451
21.1 INtrODUCtION
New trends in product development are promoting intense research on alternative methods
for food processing and preservation. Traditionally, foods have been processed and preserved by
thermal treatments that prevent food from spoilage and potential human diseases by inactivating
microorganisms. On the other hand, it also possesses some limitations such as impaired freshness,
loss of colour, and loss of heat-liable nutrients such as ascorbic acid. In view of this, non-thermal
food processing concept was born. Non-thermal processing of food, as the name suggest, means
processing food products much below the temperatures involved in conventional processing; thus,
food flavours, nutrients and other organoleptic properties undergo minimal changes. The sole prin-
ciple behind the use of this technology is to inactivate undesirable microorganisms present in the
food and also certain endogenous enzymes without affecting the nutritional and sensory compo-
nents that are normally affected during severe heat treatment. These non-thermal processes require
specialized equipment, trained personnel, and are relatively very expensive.
Foods can be non-thermally processed by (1) irradiation, (2) high hydrostatic pressure, (3)
electro-magnetic methods such as pulsed electric fields (PEFs) and oscillating magnetic fields, (4)
ultrasound, (5) micro and ultrafiltration, (6) ohmic heating and cryogenic grinding, (7) the use of
antimicrobials, bacteriocins, etc. Each of these techniques can be used either alone or in combi-
nation to enhance product quality, decrease processing time, and effective bacterial and enzyme
inactivation. Another advantage of non-thermal processes is that, in general, they utilize less energy
than thermal processes. Non-thermal processing of food products is therefore being developed as an
alternative to traditional thermal food processing procedures.
433
In this review, a range of technologies, from those that have been received negatively and those
that have the potential to be received negatively in the future, to those that have had more favour-
able societal response, are analysed, characterised, and compared. Approaching the development of
novel food technologies through investigation of psychological, socio-cultural, and historical issues
is an essential element of product commercialization. Patterns in their trajectories of societal accep-
tance are identified, from the perspective of both the technologies and consumers.
21.2 FOOD IrraDIatION
Food irradiation is a preservation method that involves exposing food to gamma rays, X-rays,
or electrons. Food irradiation technologies exploit the part of the electromagnetic spectrum that
encompasses wavelengths shorter than 10−10 m. Radiation from this region of the electromagnetic
spectrum is called ionizing radiation, because it can ionize materials and molecules that it encoun-
ters. Gamma radiation, X-ray, and e-Beam radiation are examples of ionizing radiation and radio
waves, microwaves, and ultraviolet (UV) radiation are examples of non-ionizing radiation.
Unfortunately, food irradiation is one of the least understood food processing technologies. Food
irradiation involves exposing the food to a carefully controlled amount of ionizing radiation for an
effective period of time. Irradiation makes food safer and spoilage resistant without compromis-
ing taste, texture, aroma, or nutritional values. The process destroys insects, moulds, fungi, and
pathogens that cause food-borne diseases or food spoilage. Its main benefits are that it kills patho-
genic organisms, including Salmonella, Listeria monocytogenes, Campylobacter and E. coli, whilst
otherwise having minimal impact on the quality of food products (Diehl, 1993).
Food irradiation helps in (1) reducing high food losses from infestation, contamination, and
spoilage; (2) controlling food-borne diseases; (3) reducing transmittal of diseases by international
trade; (4) lessening strict regulations and prohibition of food chemical treatments; and (5) extending
shelf life.
Gamma radiation is generated by photons emitted from radioactive isotopes such as cobalt-60
and cesium-137. These are radioactive, which poses significant challenges in acquiring, transport-
ing, storing, and safeguarding them. Because of the potential for theft of the radioactive mate-
rial, there is a strong push by the International Atomic Energy Agency (IAEA), the U.S. National
Nuclear Security Administration (NNSA), and the U.S. Defense Threat Reduction Agency (DTRA)
to limit the commercial use of these radioactive materials.
Although e-Beam and X-ray irradiation technologies employ ionizing radiation, the radiation
is not produced by radioactive materials. Rather, it is generated by specialized equipment called
industrial electron accelerators. These accelerators are switch-on/switch-off technologies that can
be turned off when not in use. In contrast, radioactive sources, such as cobalt-60 and cesium-137,
cannot be switched off and generate gamma radiation continuously. The ability to switch the radia-
tion source on and off has major implications in terms of operating costs, worker safety, and the
carbon footprint of an e-Beam or X-ray food processing facility.
Electron beam and X-ray technologies can be used to treat food and food ingredients to elimi-
nate microbial pathogens (i.e., pasteurization), or at higher doses to sterilize food ingredients. They
can also be used at very low doses for phytosanitary treatments, which eliminate insects and pests
on agricultural products to prevent their accidental introduction into other areas. The actual pen-
etration depth of electrons or X-rays also depends on the density of the target food item. Energies
from gamma, X-ray, and electron beam sources are too low to induce radioactivity in any material,
including foods. Table 21.1 shows the dose rates of gamma, X-ray, and e-Beam irradiation at differ-
ent beam energy levels, as measured at the National Center for Electron Beam Research at Texas
A&M University.
ConsuMer ACCePtAnCe AnD Future trenDs oF non-tHerMAL-ProCesseD FooDs 435
table 21.1 the Beam Energy and Corresponding Dose rate of

e-Beam, Gamma, and X-ray radiation
Source Energy, MeV Dose rate, Gy/sec
electron beam 10 ~3,000
8.5 ~3,000
Gamma 1.59 (from lanthanum-140) ~0.06–0.12
X-ray 5 ~100
0.1 ~0.01
Food processing applications of e-Beam technology can be broadly divided into low-energy
(<1 MeV), medium-energy (1–8 MeV), and high-energy (8–10 MeV) applications. Current
low-energy applications include the inline sterilization of packaging materials and the inline dis-
infestation/sterilization of seed surfaces. Medium-energy applications include phyto-sanitary
treatment of packaged fruits and vegetables. High-energy applications include pasteurization of
packaged meats, spices, seafood, and food ingredients. The FDA has established maximum doses
that can be applied to different types of foods, spices, and seasonings (Table 21.2). The USDA Food
Safety and Inspection Service (FSIS) oversees the application of this technology to fresh and frozen
meat and poultry products, and the USDA Animal and Plant Health Inspection Service (APHIS)
oversees its use for phyto-sanitary treatment of fresh fruits and vegetables.
Sensory characteristics and consumer acceptance of electron-beam irradiated commercial
samples of ready-to-eat meats (frankfurters and diced chicken) were evaluated by Johnson et al.
(2004). Samples were removed from their original packaging, repackaged in irradiation-approved
packaging, vacuum-sealed, irradiated by electron-beam at 1, 2, and 3 kGy, and stored at 4°C for up
to 32 days. Non-irradiated controls were held under similar conditions. A consumer panel evalu-
ated the effects of irradiation on the samples throughout the expected shelf life of 32 days after
irradiation. Overall acceptance, acceptance of flavour, juiciness, tenderness and mouth-feel of the
non-irradiated diced chicken and frankfurters were significantly lower than most irradiated samples
at day 18 and day 32 after irradiation, respectively. The irradiated frankfurters and diced chicken
maintained their acceptability for up to 32 and 18 days, respectively, after irradiation.
Food irradiation is supposed to be no more hazardous to achieve the same purpose than canning,
drying, heat pasteurization, or cooling process. Irradiation is a “cold pasteurization process;” it does
not raise substantially the temperature of the food being processed. Nutrient losses are small and
table 21.2 the FDa has Set Maximum allowable Dosages for Food Irradiation applications in the U.S
Food or Food Ingredient application Maximum allowable Dose, kGy
White potatoes sprouting inhibition 0.15
Fresh, non-heated processed pork Pathogen control 0.3–1.0
Wheat flour Mould control 0.5
Fresh produce insect disinfestation 1.0
Fresh produce Growth and maturation inhibition 1.0
Fresh or frozen uncooked poultry Pathogen control 3.0
products
Fresh shell eggs Pathogen control 3.0
Fresh iceberg lettuce and fresh spinach Pathogen control 4.0
refrigerated, uncooked meat products Pathogen control 4.5
(sheep, cattle, swine, and goat)
Fresh or frozen molluscan shellfish Pathogen control 5.5
Frozen, uncooked meat products Pathogen control 7.0
(sheep, cattle, swine, and goat)
often significantly similar to losses from other methods of preservation. Much of the early work on
irradiation examined foods treated at very high doses, but today, irradiation is very precise. Scientific
studies have shown that precision dose specific irradiation does not significantly reduce nutritional
quality or significantly change food taste, texture, or appearance. In September 1997, an expert group
organized by the World Health Organization (WHO), Food and Agriculture Organization (FAO),
and International Atomic Energy Agency (IAEA) concluded that doses greater than 10 kGy “will
not lead to changes in the composition of the food that, from a toxicological point of view, that would
have an adverse effect on human health.” The United States Food and Drug Administration (FDA)
has also evaluated the safety of this technology over the last 40 years and has found irradiation to be
safe under a wide range of dose levels and has approved its use for many foods.
Irradiation has a high potential and is probably one of the most versatile among the food pres-
ervation technologies. However, its development and commercialization has been hampered in the
past by unfavourable public perceptions. Acceptance of irradiation had been slowed by several
factors. Firstly, the term “irradiation” was associated with radioactivity by consumers and this is
perceived as alarming. Food irradiation is associated with “radioactivity,” which is of major public
concern. Second, the reasons for its use and the potential benefits were poorly understood by the
general public as well as health professionals.
Food irradiation is permitted in a number of countries internationally. In Europe, it is allowed
for dried herbs, spices and vegetable seasonings, and in individual countries for chicken/poultry
(as is the case in the US), and other meat products. Most research into public acceptance of food
irradiation was conducted in the 80s and 90s (Bruhn, 1998). The majority of studies were US-based,
on meat. Exceptions include studies in Brazil (Behrens et al. 2009), Turkey (Gunes and Tekin,
2006), and Europe (Sparks and Shepherd, 1994; Siegrist et al., 2006).
The research evidence suggests that people tend to be unfamiliar with food irradiation. Despite
this, they hold negative views about its application. The main consumer concerns focus on the
perceived “carcinogenicity” of irradiated foods, impaired food quality, risks to factory workers
and the environment incurred during production. Some consumers believe that the benefits accrue
differentially to manufacturers compared to consumers. The negative associations that consumers
have with food irradiation have restricted widespread implementation of the technology by industry,
as it requires significant investment in infrastructure (Henson, 1995).
Public awareness of food irradiation is low, but despite this acceptance is low. Food irradia-
tion can be characterized as a technology for which the public perceives not many personal ben-
efits. Food irradiation evokes rather negative associations, due to the name and the perception of
uncontrollability. The perceived risks associated with irradiation appear to be more varied, poten-
tially profound, and extend beyond risks of personal harm into bio-systems and bioactivity, com-
pared to those associated with the other technologies. Much of the literature suggests that public
attitudes towards food irradiation are, on the whole, negative (e.g., He et al., 2005; Gunes and Tekin,
2006; Ronteltap et al., 2007).
Consumer research can identify the questions consumers can have about a new technology.
People are primarily interested in how the new process or technology affects them. For example,
more people are more interested in how eating irradiated food affects human health than how irradi-
ated food tastes. Different driving factors determine consumers’ acceptance of novel technologies
in food industry.
The views of the public on food irradiation also appear to vary depending on the type of
food, with irradiation being perceived as more appropriate for particular foods, and unnecessary
for others. For example, research by Johnson et al. (2004) suggested that irradiation of fruit and
vegetables may be perceived as more acceptable than irradiation of meat. In the case of food
irradiation, public concerns are also linked to the perception that potential benefits are weighted
towards industry. The view that industry and producers mainly takes profit from these technolo-
gies shapes consumers’ perceptions.
For food irradiation (and possibly in the future for nanotechnology), consumer acceptance is
driven by associations with other technologies perceived to have similar characteristics, i.e., in other
words, acceptance is driven by generalised attitude towards food technologies that are perceived
to share common characteristics. This suggests that perceptions of uncontrolled use and potential
for bioactive impact beyond the original intentions of the technology proponents may be shaping
societal responses.
Consumers are often suspicious of novel food technologies, though this is not always warranted
from a scientific point of view. Food technologists and food processors should adopt a holistic
approach when introducing new technologies to reassure consumers of their safety. A simple exam-
ple of such an approach is proper nomenclature for novel technologies, which should be neither
deceptive nor frightening. Such an approach necessitates engaging everyone along the supply chain,
from the grower to the consumer, in an open dialogue to elucidate all the objectives of the novel
technologies along with their pros and cons.
High-pressure processing (HPP) technology offers the food processing industry with great oppor-
tunity to explore new product development with higher nutritional profile, good taste and texture, per-
fect aromas, and ultimately more shelf life (Nath et al., 2016). HPP uses water as a medium to transmit
pressures from 300 to 700 MPa to foods resulting in a reduction in microbial loads and thus extending
shelf life. Pressure is transmitted uniformly (in all directions simultaneously), thereby helping the
food to retain its original shape and texture even at higher pressures. HPP leads to minimal changes in
nutritional value of foods along with it helps in maintaining the “fresh”-like characteristics of foods
by eliminating degradation characteristics of high temperature usage. In comparison with thermal pro-
cessing, HPP results in foods with fresher taste, better aesthetic value, texture and nutrition.
HPP can be conducted at ambient or refrigerated temperatures, thereby eliminating thermally
induced cooked off-flavours and off-taste. This technology can prove to be a boon for heat-sensitive
food products. HPP has been extensively studied and explored by many food scientists and nutri-
tionists all over the world in paste and purees: peach puree, pumpkin puree, nectarine puree, apple
puree, strawberry puree and blackberry puree, and tomato and carrot purees.
The development of healthy foods was rated as the most important area of research by a large
majority of the companies interviewed (Katz, 2000). The method of production is also of increasing
concern to many consumers, who look for products similar to the natural ones. High pressure is a
new food processing technology developed to achieve consumer demands for fresher products with
reduced micro-biological levels and improved flavour (Vardag et al., 1995; Rosenthal and Silva,
1997; Swientek, 1999; Khamrui and Rajorhia, 2000). This non-thermal food preservation technique
could have a large food industry impact due to the benefits it can bring to products if these benefits
are well communicated to consumers. However, there is not much literature available on consumer
acceptability of this technology. There seems to be no published reports on toxicity of HPP-treated
food products although allergic reactions have been reported elsewhere (Hugas et al., 2002; Oey
et al., 2008). In general, public awareness about HPP is low. Research suggests that the public is
neutral to slightly positive about HPP, with only small cultural differences in acceptance (Butz et al.,
2003). The perceived personal and societal benefits of HPP may play a considerable role in con-
sumer acceptance (Olsen et al., 2010).
Sorenson and Henchion (2009) conducted a study on consumers’ perception on HPP ready-to-eat
chilled meals. They used soft laddering system technique to quantify the consumer perceptions.
The in-depth discussions with the consumers explored a range of issues concerning consumers’
acceptance of HPP, as well as their preferences for HPP chilled ready meal concepts. The results
of the study showed that consumers were generally receptive towards HPP of chilled ready meals.
The name of the technology also evokes mainly positive associations. Perceived risks
(personal and societal) do not seem to play a salient role in acceptance, although possible risks are
often implicitly referred to, for example, in terms of “unknown consequences,” “unnaturalness,”
“unfamiliarity,” or through negative associations with HPP or a general lack of trust in regulators
and industry. Results of research may be biased as study participants have not usually been familiar
with HPP before being enrolled in studies, only being informed of potential HPP (dis)advantages at
the outset of research.
HPP seems to be an accepted food technology. It has been introduced in various markets with-
out stirring much controversy: consumers seem to perceive benefits, without no associated risks
with this technology. Furthermore, consumers seem to associate environmental as well as work
safety benefits with this technology. For HPP food products, mainly food safety concerns rep-
resented a strong perceptual barrier to consumers’ acceptance whereas thermal processing was
positively associated with assurances of greater food safety. HPP as a means of non-thermal food
preservation might represent a greater risk in terms of food safety because “the more the food is
fully cooked the more likely the chances of bacteria being killed.” In a research conducted by
Sorenson and Henchion (2011) on understanding consumers’ cognitive structures with regard to
HPP of chilled ready meals category, it was found that consumers were generally receptive towards
the concept of HPP chilled ready meals. However, perceived food safety risks related to low aware-
ness levels of the technology, coupled with misconceptions regarding the influence of HPP on
elements of eating quality represented significant barriers to consumer acceptance. Hierarchical
value maps (HVM) revealed important insights into the motivational cognitive structures of spe-
cific consumer segments with important implications for the technical development and strategic
marketing of HPP foods.
HPP has been judged to be relatively similar to conventional process technologies in terms of
overall consumer acceptability (Butz et al., 2003; Deliza et al., 2003, 2005; Evans and Cox, 2006;
Mireaux et al., 2007). However, Cardello et al. (2007) and Lampila and Lahteenmaki (2007) tried
to explain these findings in terms of HPP being less well known, and less emotive, than technologies
such as food irradiation and GM foods.
Nielsen et al. (2009) and Cardello et al. (2007) cautioned that an element of consumer uncertainty
and risk could still be expected with HPP foods. For example, Nielsen et al. (2009) found that while
consumers were receptive towards the improved sensory and nutritional benefits associated with
HPP, they also expressed negative attitudes towards their cost as well as the lack of information in
the public domain. The provision of benefit related information is considered as a key success factor
in terms of garnering high levels of consumer acceptance for HPP foods (Butz et al., 2003; Cardello,
2003; Deliza et al., 2003).
Worryingly, for some companies there can be an expectation that the adoption of new tech-
nologies that offer specific benefits will automatically create value and lead to a distinct compet-
itive advantage in the marketplace. From a managerial perspective, it is important for companies
to understand that technological advances that seek to add value do not necessarily give rise
to consumer value. For example, HPP units are currently marketed on the basis of offering
minimally processed foods with superior nutritional and organoleptic qualities, without compro-
mising food safety and shelf-life. However, a number of consumer studies have found benefits
related to health, sensory quality, or the environment to be more acceptable to consumers than
producer-oriented benefits such as a shelf-life extension (Frewer et al., 1997; Butz et al., 2003;
Cardello et al., 2007; Nielsen et al., 2009). Consequently, new product development strategies
that seek to add “superior value” to foods should be viewed and assessed from the consumers’
perspective.
In other studies, it was found that consumers have been most receptive towards HPP when
provided with information on its potential benefits (Butz et al., 2003; Deliza et al., 2003; Nielsen
et al., 2009). In another study by Deliza et al. (2005) it was found that consumers expressed a higher
intention to purchase orange juice when the technological advantages were mentioned on the labels
than when the technology was merely identified by its name.
A study with Brazilian consumers using focus groups was carried out to investigate the impact
of the information about the use of high pressure in the fruit juice production on the consumer
(supermarket shoppers) product perception (Deliza et al., 2003). Three pineapple juice labels with
different characteristics in terms of information on nutritional, sensory, and employed technology
in the juice production were created especially for the study. The results indicated that participants
showed concern about the appearance and quality of labels in general, and revealed willingness to
taste new products. They considered themselves loyal to brand, and some of them declared that brand
would overcome price. Although price was not manipulated in this study, the majority of the par-
ticipants in all four sessions mentioned price as an important attribute during their decision-making
process. This result confirmed previous work, particularly a study by Pecher and Tregear (2001), in
which price was found to be the overwhelming factor and dictating quality perception. Furthermore,
consumers in this study pointed out that when they were not very familiar with the brand name,
they very often read the label/package looking for information. This revealed a very important issue
to food producers, since non-familiar brands could be promoted by providing consumer relevant
information about the product. Results presented by Deliza et al. (1999, 2003) demonstrated that
consumers inferred product taste from the package/label, revealing many packaging attributes which
affected product expectation and perception. Product information (e.g., nutritional, sensory, safety,
and ingredients) and technology information appeared to be important package attributes. Deliza
et al. (2003) found out that three out of four consumer groups perceived products as having higher
quality when the package label included technology information. This action was perceived as reflect-
ing a greater concern for the consumer by the food producer favouring the consumer selection of
such products. The statement “Saudavel com maissabor”—healthy with more flavour—contributed
to a more positive fruit juice perception in all interview sessions.
Simply mentioning the technology—high pressure—catches the attention, but such information
is not sufficient for the consumer acceptance. As presented in the study, some respondents con-
sidered the information negative and thus affecting product acceptance leading to a low purchase
intention. Certainly, some information explaining the meaning and advantages of high-pressure
technology may lead to a higher product satisfaction, and contributing to the market introduction
of a product that offer higher nutritional and sensory qualities (Deliza et al., 2003). Effective com-
munication between the producer and the consumer about food and nutrition relies on delivering
messages that consumers find believable and that also convinces them that making healthy food
choices is achievable (Borra and Earl, 2000). The issue of consumer benefit appears to be an impor-
tant factor in determining acceptance, and reflects earlier research findings.
HPP can enable processors to produce innovative foods with fresh-like, natural-like attributes
and natural looking colour which are all aspects valued by consumers. In this context, especially
the beverage sector has many product development opportunities since a wide range of innovative
products can be made available to modern consumers using this technology. Consumers nowadays
are more interested than ever before in nutritious, healthy and convenient foods (Sloan, 2003) which
are possible by HPP. However, appropriate labels and the information they contain will become
more important than ever before, and have to be used aiming at fully informing consumer about
the advantages and benefits that HPP can deliver. Studies pertaining to consumer preferences with
regards to HP processed products are given in Table 21.3.
table 21.3 Summary of hpp and pEF Consumer Studies
440
Independent Variable(s)
author’s Sample and Methodology (Key Determinants) Dependent Variable(s) Key Findings
butz et al. 3000 consumers in France, basic information about Perceptions of HPP HPP was acceptable to the majority of consumers in
(2003) Germany, uK. Computer- HPP in a neutral format Perceptions of France and Germany.
assisted personal interviews advantages and it was important that the product price does not
HPP food in general disadvantages exceed that to conventional products and that there is
Likelihood of buying a health benefit.
those who perceived the greatest personal advantage
from the technology were most likely to buy the
product. this group tended to include a higher
proportion of young, educated people
Cardello 88 consumers, us experiment name of the technology Actual liking and concerns Females have a significantly higher concern level for
(2003) PeF, HPP, irradiation, (e.g., PeF) all technologies; rating of concern was negatively
bacteriocins, High-voltage name+objective correlated with expected liking of products believed to
pubs, heat pasteurization, description of the be processed by the technology. expected liking
non-thermal preservation, technology ratings were positively influenced by visual exposure
pulsed light chocolate pudding name+description+benefit to the product and by a safety and benefit statement.
condition
Cardello et al. 225 potential consumers of Food type, processing or interest Preserved risks associated with the technology were
(2007) novel processing techniques is production technology the most important factor influencing interest in use.
us Conjoint costs, benefits, risks, Among the emerging technologies assessed HPP
PeF irradiation, heat endorsing agencies and produced the most positive effects.
pasteurization, ionizing energy, product information
genetic modification, cold
preservation, benefits Different
food products meat, milk,
bakery, vegetables, fruit, sauce.
Cox and 294 consumers, Australia Food technologies Willing to try novel Ftns was tested and found to have predictive validity
evans survey neophobia scale technologies for consumers’ willingness to consumers foods
(2008) HPP, Pasteurized MAP, triploidy, (Ftns): 13 items produced by novel technology furthermore,
GMo, bioactive • new food convergence validity was found between the Ftns
Framed prawns technologies are and tiss
unnecessary
• Perception of risks
• Healthy choice
• information/media
trust in science scale
(tiss)
(Continued)
table 21.3 (Continued) Summary of hpp and pEF Consumer Studies
Deliza et al. 96 consumers, brazil Conjoint information about the Consumers intention to the technologies advantages were presented on
(2005) HPP Fruit Juice technologies consume pineapple juice labels, participants then understood
• name+benefits the benefits and expressed a higher product intention
• name to purchase
• no information
Deliza et al. 41 students and housewives, information about the Awareness, attitudes the results, suggest that giving consumers information
(2003) brazil. 4 focus groups HPP fruit technology, nutrition, and opinions, behaviour, and about the technology for food production-HPP. Had a
juice sensory qualities benefits concerns towards positive impact on the perception of the product.
processed foods
Acceptance of HP
freezing
Lampila and 936 consumers in the information about Acceptance of HP Generally, attitudes towards high-pressure freezing
Lahteenmaki netherlands, belgium, spain, technology benefits freezing were neutral, even though the term was unfamiliar for
(2007) and Finland Conjoint HP most consumers, when given some information about
freezing Vegetables high-pressure freezing technology; consumers
considered applying this method as appropriate,
especially if it had advantageous consequences in
the product. the processing method itself was
considered less important than price or environmental
impact when relative importance of choice criteria
was studies.
Mireaux et al. 72 consumers, Australia 13 Perceived attributes Preference scores for the HPP orange juice was perceived as interfered with
(2007) focus groups repertory Grid products produced by it was perceived as having a better quality but was
HPP, pasteurized orange juice, novel technologies less familiar. More expensive and had more unknown
margarine, farmed prawns, consequences than pasteurized juice.
beef, yoghurt.
nielsen et al. 97 consumers, norway, Description of the Attitudes towards HPP Participant’s evaluation PeF and HPP processes
(2009) Denmark, slovenia, Hungary, technologies + benefits and PeF technologies showed that environmentally friendly and more
serbia, slovakia. 12 focus and HPP and PeF- natural products were seen as the many advantages,
groups HPP, PeF baby food treated products while they were concerned about body and health,
ConsuMer ACCePtAnCe AnD Future trenDs oF non-tHerMAL-ProCesseD FooDs
and juice the high price, the lack of information about the
technologies and a general scepticism.
(Continued)
441
442
table 21.3 (Continued) Summary of hpp and pEF Consumer Studies
olsen et al. 609 consumers, norway, information about Likelihood of choice for european consumers view HPP juice and PeF juice as
(2010) Hungary, slovakia. Conjoint • technologies novel processed apple good alternatives to pasteurized juice if the price is
PeF, HPP, pasteurized untreated • Price juice right.
Apple Juice • taste Consumers are not willing to pay a premium price for
• Consequences of use HPP or PeF juice.
• environmental some consumers emphasize health consequences,
• Health while others focus
• Values
• Hedonism
• benevolence
sonne et al. 120 consumers, norway, Description of the Attributes consequences Consumers seem to recognize and appreciate the
(2012) Denmark, Hungary, slovakia technologies+benefits and values explaining benefits that food produced with HPP and PeF have
Qualitative laddering interviews preference for novel juice to offer when this information is provided
PeF, HPP, pasteurized Apple PeF juice was the product that most consumers were
Juice ambivalent about PeF products were perceived to
have advantages but in many cases, consumers also
appeared unsure about the risk. When raced with a
choice, consumers tended to go for HPP perceived as
offering the same benefits and at the same time
carrying less risk.
21.4 pULSED ELECtrIC FIELD
High-intensity PEF is regarded as a non-thermal food preservation technology that involves

the discharge of high-voltage electric short pulses through the food product. Electric fields in the
range of 10–80 kV/cm generated by the application of short high-voltage pulses (μs) between two
electrodes causing microbial inactivation at temperatures below those used in thermal process-
ing (60°C–70°C). The precise mechanism by which microorganisms are inactivated by PEFs is by
permeabilization of microbial membranes. Unlike traditional methods of electric pasteurization of
foods, PEF utilizes electric energy in the form of short pulses resulting in mechanical effects on cell
membrane without significant heating of the food.
The effects of PEF on bio-membranes have been widely investigated since the use of PEF has
attracted great interest in several scientific areas such as cell biology, biochemistry, pharmacology,
and food technology (Zimmermann, 1994; Ho and Mittal, 1996). With the use of electric fields,
PEF technology enables inactivation of vegetative cells of bacteria and yeasts in various foods. As
bacterial spores are resistant to PEFs, applications of this technology mainly focus on food-borne
pathogens and spoilage microorganisms, especially for acidic food products.
In addition to the volumetric effect of PEF technology in controlling the microbiological safety
of foods in a fast and homogenous manner, successful application provides extended shelf life
without the use of heat to preserve the sensory and nutritional value of foods. PEF technology
has the potential to economically and efficiently improve energy usage, besides the advantage of
providing microbiologically safe and minimally processed foods. In 2005, PEF-processed fruit
juices were introduced to the U.S. market (Ravishankar et al., 2008). The PEF process is consid-
ered safe because no dangerous chemical reactions have been detected (Vega-Mercado et al., 1997;
Soliva-Fortuny et al., 2009).
PEF offers the ability to inactivate microorganisms with minimal effects on the nutritional,
flavour, and functional characteristics of food products due to the absence of heat. PEF technology is
mainly intended for preservation of palpable fluid or semi-fluid foods. In particular it could be used to
improve the shelf-life of milk (Sampedro et al., 2005), green pea soups (Vega-Mercado et al., 1996),
liquid whole eggs (Ma et al., 1997), and fruit juices (Heinz et al., 2002; Hodgins et al., 2002). The
sensory attributes of fruit and vegetable juices are reported to be well preserved with fresh like qual-
ity and extended shelf life after processing. Other PEF-processed foods include yogurt drinks, apple
sauce, and salad dressing (Bendicho et al., 2002; Hodgins et al., 2002).
As for HPP, public awareness of PEF is low and public responses to PEF vary from slightly posi-
tive to slightly negative (Cardello et al., 2007). Consumer concerns about PEF are not considered
significant, although it is perceived to have relatively few benefits (Nielsen et al., 2009). PEF has
more negative associations than HPP (for example, linked to electricity, irradiation, and microwaves)
(Sonne et al., 2009). As for HPP, a bias may arise, given that participants have responded after the
provision of information about PEF.
For both HPP and PEF, there is little evidence to suggest high levels of consumer negativity.
Whilst this may reflect fewer studies being conducted and hence reduced availability of relevant
data, this in turn may reflect lack of societal controversy. Despite this, some consumer concerns
have been identified (Table 21.3). In this respect, the least controversial technologies (HPP and
PEF) were both highly contained and potentially limited in both time and space in terms of their
perceived impact, in particular in terms of bioactive effects.
In other words, while food is bio-actively altered in HPP and PEF, the technologies themselves
are not considered to be “bioactive” in a way that may impact on current and future generations of
humans, animals, and plants. This also suggests that perceived “unnaturalness” of a technology
in itself does not reduce consumer acceptance, but that perceptions of “uncontained bioactivity”
associated with technological application may result in rejection. In addition, the issue of consumer
choice, traceability or personal control over exposure appears to be a key issue for the least accepted
technologies. Facilitating consumer choice through efficacious labelling may support an acceptable
compromise for more controversial technologies.
PEF and HPP are also not well known by the public, although the evidence available sug-
gests that they are of potentially lesser concern, PEF and HPP are also associated with far fewer
potential applications than, other technologies (simplifying regulation and containment of use), and
are unlikely to be perceived as bioactive. As noted, their application is relatively easily contained.
Guo et al. (2014) reported regarding microbial safety, bioactive compounds, physico-chemical
properties, and consumer acceptance of PEF-processed pomegranate juice and found that consumer
acceptance for PEF-treated pomegranate juice sample was similar to the unprocessed juice sample.
This study also demonstrated that PEF technology extended microbial shelf-life and preserved the
major quality and nutritional characteristics of pomegranate juice.
21.5 COLD pLaSMa tEChNOLOGY
Plasma is a term which refers to the fully ionized gas composed of various substances such
as photons (light in nature) and free electrons, along with atoms (heavy in nature) in excited state
having a neutral charge. It is regarded as the fourth state of matter which starts from solid to liquid
state, liquid to gas and finally to plasma (Misra et al., 2011). Plasma is generated when an inert
gas comes in contact with electricity then reactive substance composed of charged particles, free
radicals, photons, and various radiations are formed. This whole formulation is known as plasma.
Plasma produced at an ambient temperature is referred to as cold plasma having a temperature of
30°C–60°C, which is mostly preferably used in the food processing industries (Misra et al., 2011).
Cold plasma is an emerging technology in the recent era and is gaining fame for its unique char-
acteristics like application at low or ambient temperature which helps in retaining the integrity
and quality of food products. Cold plasma has proved to be efficient in sanitizing equipment for
inactivating the food-borne pathogens from fresh produce and packaging materials. It also helps in
catalysing certain manufacturing processes, acts as an active packaging, and retards browning reac-
tion in fruits and vegetables. Being a cold treatment it is effective in retaining the texture, sensory
and functional properties of foods. Cold plasma withholds the ability to manufacture high-quality
fresh and processed food products. The non-thermal or cold plasma technology is now the prime
consideration in food processing industries especially packaging sector.
In general plasma is classified into two types: thermal and non-thermal. Thermal plasma gen-
eration requires high-pressure and temperature with heavy electrons. Non-thermal or near ambient
temperature plasma (NTP) is generated under atmospheric or vacuum at temperatures of 30°C–60°C
requiring low energy. NTP can be formed by electrifying or using electromagnetic waves on gas at
reduce pressure, having a thermodynamically non-equilibrium nature.
There are several approaches in creation of plasma at atmospheric pressure, such as dielectric
barrier discharge, corona discharge and gliding arc discharge (AD), which are of interest in food
industries and require mild conditions (Misra et al., 2011). The accumulation of charged reactive
particles on food through pressure or dynamic shock can rupture the cell membrane of microbes.
Oxidation of the lipids, amino acids and nucleic acids with reactive oxygen species and nitrogen
species cause changes that lead to microbial death or injury.
Cold plasma technology is successfully studied for effective microbial inactivation of
Escherichia coli from fresh produce (Sharma, 2009; Bermudez-Aguirre, 2013), Aspergillus para-
siticus and Penicillium sp. from seeds of various vegetable, legumes, and cereals (Selcuk, 2008),
Erwinia carotovora in potatoes (Mireaux, 2007), Listeria monocytogenes from plastic trays, paper
cups, and aluminium foil, etc. Other than decontamination, plasma technology works in various
other phenomena such as effect on seed germination (Sera, 2012) and retarding browning reaction
(Tappi, 2016).
The packaging process is an important critical control point in a hazard analysis and critical
control point (HACCP) system (Mittendorfer et al., 2002) so most regulatory guidelines specify
microbiological requirements for food packaging materials. Food packaging materials are intended
to preserve food quality along the distribution and storage chain and also to protect it from deterio-
ration, damage or outside contamination. If food packaging is not properly sterilized this may cause
further contamination of the food from the packaging surface and consequently lead to health risks
and economic losses (Misra et al., 2011). Cold plasma sterilization is a chemical free, fast and safe
approach, applicable to a wide range of packaging materials and does not result in any residues.
However, its adoption for mass-production in the food packaging sector is limited by the treatment
times, which often extend to minutes; extended sterilization periods are not affordable by the food
industry. Combining the CP treatment with other non-thermal processes could be a possible future
breakthrough in this field. In this case, synergistic effects may be more considerable; however,
scaling up this technology remains for the moment a challenge to be solved.
21.6 CONSUMEr rESpONSES tO rECENt INNOVatIONS

IN FOOD prOCESSING tEChNOLOGIES
Food has long been associated with numerous food-borne diseases so consumers nowadays have
increasingly become aware of healthy, hygienic habits, and diet. Microorganisms and pathogens
that cause spoilage are a major problem concerning the food processing industries and regulatory
bodies, as they have an unfavourable impact on the health and economy of the public.
To control and eliminate pathogenic microorganisms effectively, sanitation techniques are used
for post-harvest. In a study, concern levels for 20 different traditional and novel food technologies
and treatments were obtained from a group of U.S. consumers (Cardello, 2003). Among the food
technologies/treatments that evoked the greatest concern were genetic engineering (rank = 1), the
addition of bacteriocins (2), irradiation (3) and pulsed X-rays (4). Of somewhat lesser concern were
such technologies/treatments as UV-light (6), PEFs (8), and oscillating magnetic fields (11). Other
innovative and emerging technologies evoked still lower concern, e.g., hydrostatic pressure (14),
radiofrequency heating (15) and electrical resistance heating (17), while those evoking the least
concern were the traditional processes of “thermal energy” (19) and “heat pasteurization” (20).
Though non-thermal techniques for food processing possess numerous advantages, the main
drawback is low consumer acceptability owing to costlier equipment, harmful rays, disease-causing
microorganisms, effects of electro-magnetic fields, use of foreign material like bacteriocins, etc.
Consumer acceptance is one of the major barriers to widespread adoption of non-thermal-processed
foods (Molins et al., 2001). There has already been an increased negative perception towards pro-
cessed foods and a number of societal factors have contributed to this, including diminishing
appreciation of scientific contributions, as well as lack of familiarity with farming system among
urban-based consumers.
Lack of knowledge among consumers regarding innovative and emerging food technologies can
serve as a major impediment to their acceptance. Thus, effective communication regarding details
of the technologies and their benefits become essential for successful marketing of these products.
Several investigators have examined the role of product and benefit information on the acceptance,
likely purchase and use of foods processed by novel technologies (Pohlman et al., 1994; Bruhn, 1995;
Hashim et al., 1996; Schutz and Cardello, 1997; Jaeger et al., 2004; Lusk et al., 2004). In these stud-
ies, the information has often focused on the benefits to be achieved from the new technology and on
endorsements of the technology and its benefits by scientific, governmental, or other food industry
organizations (Frewer et al., 1997). In such studies the information is made available to the consumer
either on product labels, on in-store displays, through pamphlets, manufacturer websites, or other forms
of media marketing. In a study on the acceptance and purchase potential of irradiated pork sandwich as
compared to non-irradiated, irradiated one which received both types of information (benefits and risk)
though, the purchase bids also decreased and attitudes became more negative. In fact, the effect was
virtually the same as that of receiving the negative information alone (Hayes et al., 2002).
In addition to product information, product exposure and trial (tasting) also has been shown
to reduce the reluctance of consumers to choose or purchase foods processed by novel techniques
(Schutz et al., 1989; Pohlman et al., 1994; Frewer et al., 1996) or containing novel ingredients
(Luckow et al., 2005).
Dozens of studies have been carried out by scholars to investigate the acceptance of irra-
diation technology (Henson, 1995; Hayes et al., 2002; Nayga et al., 2004). Cardello (2003) and
Mireaux et al. (2007) explored the technologies by thorough study specifically on high-pressure
and/or pulsed-electric field treatment (Butz et al., 2003; Deliza et al., 2003, 2005; Lampila and
Lahteenmaki, 2007; Nielsen et al., 2009; Sonne et al., 2009; Olsen et al., 2010). Although additional
research is needed to optimize micro-biological and enzymatic inactivation in order to ensure shelf-
stable products, non-thermal technology offers unique opportunities and some challenges to the
food industry (Sizer et al., 2002).
Consumers are becoming increasingly concerned about both the quality and safety of food
they eat. These quality and health-related perceptions mean that, for consumers, it is essential
that they associate processed products with adequate (and technologically appropriate) hygiene
practices. For example, food irradiation has been rejected by European consumers, despite reduc-
tions in micro-biological risk. Cardello et al. (2007) suggested that using different terminology
could influence people’s attitudes: their research found that people were less concerned about
“ionizing energy” than about “irradiation” (bearing in mind they may not have known that the two
terms have the same meaning in practice).
A food which meets nutritional requirements is unlikely to be accepted by consumers if they
do not like the flavour or other quality attributes. Despite the mentioned importance of the sen-
sory properties of the product, studies examining consumer attitudes towards new technologies
used in food production have shown that consumers are also becoming increasingly interested in
non-sensory food qualities. Aspects such as nutritional quality, microbiology safety, agro-chemical
residue and environmental pollution are all examples of consumer concern (Frewer et al., 1998).
Within this context, the use of technologies harmless to the environment may contribute to per-
ceptions of increased consumer benefits and satisfaction. However, the negative impact of new
technology and/or food processing may be a source of consumer concern (Deliza et al., 1999).
Therefore, important issues to be investigated are consumer’s opinions, beliefs, and attitudes
towards the use of non-conventional technologies in food processing.
Advantages in terms of health, environment and safety have been shown to lead to more posi-
tive consumer reactions when they are linked to HPP- and PEF-treated products (Butz et al., 2003;
Mireaux et al., 2007; Nielsen et al., 2009; Sonne et al., 2009; Olsen et al., 2010). HPP and PEF
treatment improves a food products’ nutritional value, compared to conventional heat treatment,
is perceived by consumers to be an important benefit. So is also the gastronomic quality of the
product. An improved fresher taste is stated as an advantage. In addition, consumers perceive PEF
and HPP processes to use less energy and to be more environmentally friendly than conventional
processes (Cardello et al., 2007). In addition to this, consumers perceive costs to be linked to HPP
and PEF-treated products. Consumers seem to be concerned that HPP and PEF processes result in
more expensive products and generally, consumers are not willing to pay a premium for HPP and
PEF-treated products (Butz et al., 2003; Olsen et al., 2010).
The findings indicate that HPP products are easier to accept than PEF products (Butz et al., 2003;
Cardello et al., 2007; Mireaux et al., 2007; Nielsen et al., 2009; Sonne et al., 2009). Consumers’ dif-
ficulties in imagining and understanding the PEF technology may result in a stronger influence from
their general trust in new technologies. In this respect, PEF technologies may lean more towards
technologies like genetic modification and irradiation.
PEF products were perceived to have advantages, but in many cases consumers also appeared
unsure about the risk. Consumers were sceptical towards PEF because of its name: pulsed electric
field. This scepticism is partly due to the fact that the name generates a fear of electricity. When
faced with a choice, consumers tended to go for HPP juice which was perceived as offering the same
benefits at the same time carrying less risk. The importance of consumers perceiving the advan-
tages of HPP and PEF products was also found in a European study including 3000 adults (Butz
et al., 2003). The study found that 67% of the participants accepted HPP concluding that consum-
ers are prepared to buy HPP products that have advantages, but not disadvantages. Cardello (2003)
also found expected liking ratings to be positively influenced by visual exposure to the product and
benefit statements.
As for technologies in general, consumers of HPP and PEF are inclined to be more positive
towards technologies they understand (Lampila and Lahteenmaki, 2007). Hence, experience with
the products and more information about the technologies seems to be the key to achieve consumer
acceptance of products manufactured by means of these new technologies. Food producers and food
scientists must provide the evidence that will convince consumers that this technology is safe to use
in connection with food processing.
In a study conducted by Lee et al. (2014) on consumer acceptance of HPP, PEF, and heat-treated
beverages by Chinese consumers based upon the type of information provided, it was observed
that type of information had drastic effect on the liking and acceptance of the technology applied.
Provision of detailed information about the mentioned technologies changed their purchase attitude
towards HPP and PEF. Chinese consumers liked these technologies because of fewer additives
mentioned on the labels.
Processing technologies aiming to enhance food quality and safety are widely applied in the
food chain. It has been found that consumers’ product preferences have now become more depen-
dent on process characteristics (Grunert, 2005). Although consumers may not always understand
the technical issues, they often report preferences for particular practices, such as organic produc-
tion, minimal and green processing (Guerrero et al., 2009), while disliking others such as genetic
modification (Da Costa et al., 2000; Nielsen et al., 2009; De Barcellos et al., 2010), irradiation
(Morkbak et al., 2012), microbial decontamination (Korzen et al., 2011), or marination by injection
(De Barcellos et al., 2010).
The acceptance of a technology depends on the consumer’s perception of benefits and associ-
ated risks. These include the impact of the technology on taste, convenience, nutritional value, the
perceived safety of the process or technology, the magnitude of the risk the technology reduces, and
the effect of the technology on the environment (Table 21.4).
Public acceptance is influenced by perceived credibility of data in public domain, and dem-
onstrated responsibility of industry. Trust in regulatory policy and unbiasedness of regulators
are also important food chain factors. Consumer attitudes towards food technologies are also
strongly embedded in pre-existing fundamental attitude structures (Scholderer and Frewer,
2003). Consumer acceptance or rejection of food processing technologies also depends on the
amount of information that is provided, as was illustrated by Deliza et al. (2003) and Cardello
(2003). Proponents of particular technologies often assume that negative consumer attitudes can
be changed by providing more information to correct the so-called “knowledge deficit,” i.e., to
overcome rejection of a technology solely due to simple unawareness (Hilgartner, 1990; Teisl
et al., 2009). However, several studies have shown that simple information provision does not
guarantee more positive attitudes (Rollin et al., 2011). Information can activate existing fears
and concerns about food technologies (Cox et al., 2007) and even lead to boomerang effects
(Scholderer and Frewer, 2003).
Specifically, the provision of information about tangible benefits is considered a key factor in
shaping consumer acceptance of food technologies. Positive framing of technology information
might enhance consumer acceptance. Several studies showed that consumer-oriented attributes
table 21.4 a Summary of the Key Factors related to public acceptance of three Novel Food
technologies
Issues Impacting on high-pressure processing pulsed Electric Field
acceptability Food Irradiation (hpp) processing (pEF)
Perceived personal Perceived personal Perceived benefits relate to Perceived benefits relate
benefits (Health, benefits do not drive those envisaged by the to those envisaged by
economic, social, acceptability, though proponents of the technology proponents
environmental) recent experiences with technology and primarily similar to HPP.
food-home illness are relate to better food safety
associated with and quality with extended
increased acceptance. shelf life.
However, many
consumers do not know
what irradiation is.
Perceived societal some research Perceived environmental Perceived benefits relate
benefits (Health, indicates that people benefits are associated as environmentally
economic, social, think beef irradiation is with environmental impact. friendly with less waste.
environmental) unnecessary. environmental impact of environmental impact of
process and worker safety process was mentioned
issues also mentioned as as influential in
influential in positively evaluation PeF.
evaluating HPP
Differential accruement Consumers see the nLA nLA
of risks and benefits benefits of irradiation
(Fairness) mostly accruing to
manufacturers/retailers.
ethical concerns nLA reported to have no impact. nLA
Perceived personal Consumer’s health Perceived risks rare and not Perceived risks rare and
risks (Health, concerns (e.g., high, related to safety and not high, related to
economic, social, Carcinogenicity of allergen city. safety and allergen city
environmental) irradiated food
products) and impaired
food quality.
Perceived societal risks Concerns focus on the nLA nLA
(Health, economic, risks to factory workers
social, environmental) and environment (e.g.,
escape of radiation
from irradiation
facilities and risks
associated with the
transportation of
radioactive
Perceived efficacy of nLA Attitude towards food Attitude towards food
regulatory framework producers (positive and producers (negative)
negative) was mentioned. was mentioned.
institution that evaluate institution that evaluate
process safety influential in process safety
consumer evolution of influential in consumer
HPP. evolution of PeF.
Cognitive associations/ Cognitive associations name of technology mainly name of technology
attitude activation appear primarily linked positive for HPP, though mainly negative
to the concept of some activation of association- similar to
radioactivity and/or attitudes related to irradiation and
radioactive irradiation and GMo. microwave ovens with
contamination. some some fear of electricity
consumers do not or electrical impulses
make these and their
associations. consequences.
(Continued)
table 21.4 (Continued) a Summary of the Key Factors related to public acceptance of three Novel
Food technologies
Issues Impacting on high-pressure processing pulsed Electric Field

acceptability Food Irradiation (hpp) processing (pEF)
Public awareness Public awareness is not Awareness low. Awareness low.
(Familiarity) high. there is evidence
to suggest that many
consumers are
ambivalent.
Perceived scientific nLA HPP orange juice was Perceptions of unknown
knowledge/uncertainty perceived as having long term
unknown consequences. consequences.
Perceived naturalness nLA naturalness/unnaturalness naturalness/
often mentioned regarding unnaturalness often
HPP. mentioned regarding
PeF.
Controllability/choice there is some evidence nLA nLA
(labelling/traceability) to suggest that
consumers prefer
labelled products.
Level of consumer/ nLA nLA nLA
public involvement in
technology/product
development
trust in science and nLA scepticism/mistrust has scepticism/mistrust has
regulation been found. been found.
socio-cultural overall, consistent no difference in acceptance east european
differences differences have not between finish, Dutch/ respondents seemed
(socio-economic, been found specifically belgian and spanish more concerned about
demographic, cultural) for food irradiation, samples. Willingness to by electrical impulses than
there seems to be a HPP products varied northern european
group of people that between German, French responders. Women
are sceptical about it and uK samples. no more concerned about
are generally sceptical salient differences PeF than men.
about it are generally between e. european and
sceptical about n. european samples
scientific and women more concerned
technological about HPP than men.
innovation.
nLA—no literature available.
such as nutrition or taste are more acceptable to consumers than producer- or industry-oriented
benefits such as extended shelf life (Sorenson and Henchion, 2011), or indirect and intangible ben-
efits such as environmental gains (Cox et al., 2007). Concern about a technology influenced by
flavour expectations, (Lahteenmaki et al., 2002; Cardello, 2003). Researchers found that flavour
ratings were lower when people were told the product was produced by a new processing method.
Information about the production process might not only influence consumer expectations, but also
the taste (liking) of a food product (Caporale and Monteleone, 2004). Flavour ratings increase when
people see the produce processed by the new technology, when statements about safety are made,
and when benefits are described (Schutz et al., 1989; Bruhn, 1995; Frever et al., 1997; Cardello,
2003). Consistent with these findings, Tuorila et al. (1994) found that uncertainty associated with
a novel product may introduce a degree of expected disliking, but additional factual information
reduces this uncertainty and improves expected liking.
The people tend to have confidence in foods they perceive to be natural, in contrast to the sus-
picious attitudes they often adopt towards foods processed using novel technologies. Furthermore,
familiar and “natural” technologies are more easily accepted by consumers (Lampila and
Lahteenmaki, 2007). For example, Iaccarino et al. (2006) reported that industrially processed
soppressata salami received lower expectation scores than traditionally made soppressata. Cox
et al. (2007) in Australia found that their respondents perceived triploidy in prawns more posi-
tively than prawns treated with electron beams or irradiation, because triploidy was considered
more “natural” than the two treatment technologies.
A large body of research has examined the factors influencing consumer acceptance of new pro-
cess technologies. These factors include: trade-offs between perceived benefits and risks (Cardello
et al., 2007; Lampila and Lahteenmaki, 2007; Christoph et al., 2008); technology neophobia
(Cox and Evans, 2008); unforeseen social and moral concerns in relation to long-term effects of
new technologies on human health (Frewer et al., 1998; Onyango et al., 2006); perceived threat to
the food chain and the environment (Grunert et al., 2001); consumer characteristics such as cultural,
psychosocial, and lifestyle factors (O’Connor et al., 2005; Evans and Cox, 2006); product-oriented
factors like product experience and credence dimensions of quality (Grunert et al., 2001; Cardello,
2003; Scholderer and Frewer, 2003); and trust in food industry and regulatory bodies (Grunert et al.,
2001; Lassen et al., 2002).
In-depth consumer research (for example, using a range of psycho-social measures and con-
sumer research methodologies) is needed to develop predictive models of consumer acceptance of
novel food technologies. In particular, consideration of how consumer acceptance varies between
different individuals, socio-political contexts, and cultures, and their interactions with technology
characteristics, may be needed for technologies that are associated with potentially controversial
characteristics.
There is also some (less disputed) evidence which suggests that people with higher education
qualifications (Frenzen et al., 2001; Gunes and Tekin, 2006) and people on higher incomes (Frenzen
et al., 2001; Rimal et al., 2004) have more positive attitudes towards food irradiation. Again, how-
ever, He et al. (2005) have come to different conclusions; for example, they found evidence for more
negative attitudes among the former group.
Differences in risk and benefit perceptions may shape consumer reactions to specific appli-
cations. Research on risk perception has tended to focus on high-profile and dramatic potential
hazards at the expense of familiar ones (Hawkes and Rowe, 2008). However, other factors. such
as affective response, attitudes to technology overall, prioritization of environmental conservation,
and trust in governments and industry actors may be influential. A greater number of “degrees of
freedom” in terms of diversity of application appear to signal increased uncontrollability (both by
science and those exposed).
21.7 CONCLUSIONS
This chapter has indicated a number of factors that are associated with consumer responses
towards food technologies. Modern insights of consumer behaviour show that the picture is not
as straightforward. Implicit, often unconscious, gut feelings and the influence of the environment
are also influencing consumer choice even without the awareness of the consumers themselves.
Although the mechanisms behind consumer acceptance of major novel technologies are the same
as for technologies in general, we observe differences when moving down from the abstract level
to technology-specific level. If consumers can control consumption of associated products then it is
anticipated that consumer acceptance is likely to be higher compared to situations where applica-
tions are uncontained (in particular in terms of environmental release) and untraceable. Societal
concerns are associated with the taste and “bioactive” characteristics of technologies, and their
potential for impact on biological systems. In the case of the latter, simultaneous proactive and
engaged risk assessment which is potential determinant of consumer acceptance is needed, together
with longitudinal analysis of how consumer responses are shaped by experience with products and
regulatory policies. The chapter appears to suggest that there are research papers published which
utilise hypothesis-driven research methodologies rather survey-based methodologies. Technologies
evolution for preparation of the functional food, being characterised as being “bioactive,” raise
some concern related to unpredictable effects and uncontrolled use, as well as some ethical concern
rEFErENCES
Barbosa-Canovas, G. V., Tapia, M. S., Cano, M. P. (2005). Novel Food Processing Technologies. Boca Raton,
FL: Marcel Dekker/CRC Press.
Behrens, J. H., Barcellos, M. N., Frewer, L. J., Nunes, T. P., Landgraf, M. (2009). Brazilian consumer views on
food irradiation. Inn. Food Sci. Emerg. Tech. 10: 383–398.
Bendicho, S., Estela, C., Giner, J., Barbosa-Canovas, G. V., Martin, O. (2002). Effects of high intensity pulsed
electric field and thermal treatments on a lipase from Pseudomonas fluorescens. J. Dairy Sci. 85: 19–27.
Bermudez-Aguirre, D., Wemlinger, E., Pedrow, P., Barbosa-Cánovas, G., Garcia-Perez, M. (2013). Effect of
atmospheric pressure cold plasma (APCP) on the inactivation of Escherichia coli in fresh produce. Food
Ctrl. 34: 149–157.
Borra, S. T., Earl, R. (2000). The art and science of health promotion. Nutrn. Today. 35: 47–52.
Bruhn, A. V. A., Galvez, F. C. F., Fletcher, S. M., Misra, S. K. (1995). Consumer attitudes toward irradiated
food: Results of a new study. J. Food Protn. 58: 193–196.
Bruhn, C. M. (1998). Consumer acceptance of irradiated food: Theory and reality. Rad. Phy. Chem. 52:
129–133.
Butz, P., Needs, E. C., Baron, A., Bayer, O., Geisel, B., Gupta, B. (2003). Consumer attitudes to high pressure
food processing. J. Food Agri. Env. 1: 30–34.
Caporale, G., Monteleone, E. (2004). Influence of information about manufacturing process on beer accept-
ability. Food Quality Pref. 15: 271–278.
Cardello, A. V. (2003). Consumer concerns and expectations about novel food processing technologies: Effects
on product liking. Appetite. 40(3): 217–233.
Cardello, A. V., Schutz, H. G., Lesher, L. L. (2007). Consumer perceptions of foods processed by innovative
and emerging technologies: A conjoint analytic study. Inn. Food Sci. Emerg. Tech. 8(1): 73–83.
Christoph, I. B., Bruhn, M., Roosen, J. (2008). Knowledge, attitudes towards and acceptability of genetic
modification in Germany. Appetite. 51(1): 58−68.
Cox, D. N., Evans, G. (2008). Construction and validation of a psychometric scale to measure consumers’
fears of novel food technologies: The food technology neophobia scale. Food Quality Pref. 19: 704–710.
Cox, D. N., Evans, G., Lease, H. J. (2007). The influence of information and beliefs about technology on the
acceptance of novel food technologies: A conjoint study of farmed prawn concepts. Food Quality Pref.
18: 813–823.
Da Costa, M. C., Deliza, R., Rosenthal, A., Hedderley, D., Frewer, L. (2000). Non conventional technologies
and impact on consumer behaviour. Trends Food Sci. Tech. 11: 188–193.
De Barcellos, M. D., Kügler, J. O., Grunert, K. G., Van Wezemael, L., Perez-Cueto, F. J. A., Ueland, Q. (2010).
European consumers’ acceptance of beef processing technologies: A focus group study. Inn. Food Sci.
Emerg. Tech. 11(4): 721–732.
Deliza, R., MacFie, H. J. H., Hedderley, D. (1999). An investigation on the package features affecting con-
sumer perception of fruit juice using Repertory Grid and Focus Group methods. Brazil J. Food Tech.
2: 63–71.
Deliza, R., Rosenthal, A., Abadio, F. B. D., Carlos, H. O., Castillo, C. (2005). Application of high pressure
technology in the fruit juice processing: Benefits perceived by consumers. J. Food Eng. 67: 241–246.
Deliza, R., Rosenthal, A., Silva, A. L. S. (2003).Consumer attitude towards information on non-conventional
technology. Trends Food Sci. Tech. 14: 43–49.
Diehl, J. F. (1993). Will irradiation enhance or reduce food safety? Food Policy. 18: 143–151.
Evans, G., Cox, D. N. (2006). Australian consumers’ antecedents of attitudes towards food produced by novel
technologies. British Food J. 108(11): 916–930.
Frenzen, P. D., De Bess, E. E., Hechemy, K. E., Kassenborg, H., Kennedy, M., McCombs, K. (2001). Consumer
acceptance of irradiated meat and poultry in the United States. J. Food Protn. 64: 2020–2026.
Frewer, L. J., Bergmann, K., Brennan, M., Lion, R., Meertens, R., Rowe, G. (2011). Consumer response to
novel agri-food technologies: Implications for predicting consumer acceptance of emerging food tech-
nologies. Trends Food Sci. Tech. 22: 442–456.
Frewer, L. J., Howard, C., Hedderley, D., Shepherd, R. (1996). What determines trust in information about
food-related risks? Underlying psychological constructs. Risk Anal. 16: 473–486.
Frewer, L. J., Howard, C., Hedderley, D., Shepherd, R. (1997). Consumer attitudes toward different food-processing
technologies used in cheese production—The influence of consumer benefit. Food Quality Pref. 8: 271–280.
Frewer, L., Howard, C., Shepherd, R. (1998). Understanding public attitudes to technology. J. Risk Percep.
1(3): 221–235.
Grunert, K. G. (2005). Food quality and safety: Consumer perception and demand. Euro. Rev. Agri. Econ.
32(3): 369–391.
Grunert, K. G., Lahteenmaki, L., Nielsen, N. A., Poulsen, J. B., Ueland, O., Astrom, A. (2001). Consumer
perceptions of food products involving genetic modification: Results from a qualitative study in four
Nordic countries. Food Quality Pref. 12: 527−542.
Guerrero, L., Guardia, M. D., Xicola, J., Verbeke, W., Vanhonacker, F. (2009). Consumer driven definition
of traditional food products and innovation in traditional foods: A qualitative cross-cultural study.
Appetite. 52: 345–354.
Gunes, G., Tekin, M. D. (2006). Consumer awareness and acceptance of irradiated foods: Results of a survey
conducted on Turkish consumers. Food Sci. Tech. 39: 443–444.
Guo, M., Jin, T. Z., Geveke, D. J. (2014). Evaluation of microbial stability, bioactive compounds, physico-
chemical properties, and consumer acceptance of pomegranate juice processed in a commercial scale
pulsed electric field system. Food Bioproc. Tech. 7: 2112. doi:10.1007/s11947-013-1185-6.
Hashim, I. B., Resurreccion, A. V. A., McWatters, K. H. (1996). Consumer attitudes towards irradiated poul-
try. Food Tech. 50: 77–80.
Hawkes, G., Rowe, G. (2008). A characterization of the methodology of qualitative research on the nature of
perceived risk: Trends and omissions. J. Risk Res. 11: 617–643.
Hayes, D. J., Fox, J. A., Shogren, J. F. (2002). Experts and activists: How information affects the demand for
irradiation. Food Policy. 27: 185–193.
He, S., Fletcher, S., Rimal, A. (2005). Unwillingness to consume irradiated beef. J. Food Dist. Res. 36: 71–78.
Heinz, V., Toepfl, S., Knorr, D. (2002). Impact of temperature on lethality and energy efficiency of apple juice
pasteurization by pulsed electric fields treatment. Inn. Food Sci. Emerg. Tech. 4: 167–175.
Henson, S. (1995). Demand-side constraints on the introduction of new food technologies: The case of food
irradiation. Food Policy. 20: 111–127.
Hilgartner, S. (1990). The dominant view of popularization: Conceptual problems, political uses. Social
Studies Sci. 20: 519–539.
Ho, S. Y., Mittal, G. S. (1996). Electroporation of cell membranes: A review. Critical Rev. Biotech. 16(4): 349–362.
Hodgins, A., Mittal, G., Griffiths, M. (2002). Pasteurization of fresh orange juice using low-energy pulsed
electrical field. J. Food Sci. 67: 2294–2299.
Hugas, M., Garriga, M., Montfort, J. M. (2002). New mild technologies in meat processing: High pressure as
a model technology. Meat Sci. 62: 359–371.
Iaccarino, T., Di Monaco, R., Mincione, A., Cavella, S., Masi, P. (2006). Influence of information on origin and
technology on the consumer response: The case of soppressata salami. Food Quality Pref. 17: 76–84.
Jaeger, S. R., Lusk, J. L., House, L. O., Valli, C., Moore, M., Morrow, B. (2004). The use of non-hypothetical
experimental markets for measuring the acceptance of genetically modified foods. Food Quality Pref.
15: 701−714.
Johnson, A. M., Reynolds, A. E., Chen, J., Resurreccion, A. V. A. (2004). Consumer acceptance of electron-
beam irradiated ready-to-eat poultry meats. J. Food Proc. Pres. 28: 302–319.
Katz, F. (2000). Research priorities more toward healthy and safe. Food Technol. 54: 42–44.
Khamrui, K., Rajorhia, G. S. (2000). Non-thermal technologies for food preservation. Ind. Dairyman. 52: 21–27.
Korzen, S., Sandøe, P., Lassen, J. (2011). Don’t wash my meat: Public perceptions of decontamination in meat
production. British Food J. 113: 598–612.
Lahteenmaki, L., Grunert, K., Ueland, O., Astrom, A., Arvola, A., Bech-Larsen, T. (2002). Acceptability of
genetically modified cheese presented as real product alternative. Food Quality Pref. 13: 523−533.
Lampila, P., Lahteenmaki, L. (2007). Consumers’ attitudes towards high pressure freezing of foods. British
Food J. 109: 838–851.
Lassen, J., Madsen, K. H., Sandøe, P. (2002). Ethics and genetic engineering: Lessons to be learned from
genetically modified foods. Biopro. Bios. Eng. 24: 263–271.
Lee, P. Y., Lusk, K., Mirosa, M., Oey, I. (2014). The role of personal values in Chinese consumers’ food con-
sumption decisions: A case study of healthy drinks. Appetite. 73: 95–104.
Luckow, T., Sheehan, V., Delahunty, C., Fitzgerald, G. (2005). Determining the odor and flavor characteristics
of probiotic, health-promoting ingredients and the effects of repeated exposure on consumer accep-
tance. J. Food Sci. 70: S53−S59.
Lusk, J. L., House, L. O., Valli, C., Jaeger, S. R., Moore, M., Morrow, B. (2004). Effect of information about
benefits of biotechnology on consumer acceptance of genetically modified food: Evidence from experi-
mental auctions in United States. Euro. Rev. Agri. Econ. 31: 179−204.
Ma, L., Chang. F. J., Barbosa-Canovas, G.V. (1997). Inactivation of E. coli in Liquid Whole Egg Using Pulsed
Electric Fields Technology. New Frontiers in Food Engineering. Proceedings of the Fifth Conference of
Food Engineering, New York, USA, 216–221.
Mireaux, M., Cox, D. N., Cotton, A., Evans, G. (2007). An adaptation of repertory grid methodology to evalu-
ate Australian consumers’ perceptions of food products produced by novel technologies. Food Quality
Pref. 18: 834–848.
Misra, N. N., Tiwari, B. K., Raghavarao, K., Cullen, P. J. (2011). Non-thermal plasma inactivation of food-
borne pathogens. Food Eng. Rev. 3: 159–170.
Mittendorfer, J., Bierbaumer, H., Gratzl, F., Kellauer, E. (2002). Decontamination of food packaging using
electron beam status and prospects. Rad. Phy. Chem. 63: 833–836.
Molins, R. A., Motarjemi, Y., Kaferstein, F. K. (2001). Irradiation: A critical control point in ensuring the
microbiological safety of raw foods. Food Contr. 12: 347–356.
Morkbak, M. R., Christensen T., Gyrd-Hansen, D. (2012). Context dependency and consumer acceptance of
risk reducing strategies: A choice experiment study on Salmonella risks in pork. Food Res. Int. 45(2):
1149–1157.
Nath, P., Kale, S. J., Chauhan, O. P. (2016). High pressure processing induced changes in bioactive com-
pounds, antioxidant activity, microbial safety and color attributes of coriander paste. Agric. Res. 5: 182.
doi:10.1007/s40003-015-02008.
Nayga, R. M., Poghosyan, A., Nichols, J. P. (2004). Will consumers accept irradiated food products? Int. J.
Cons. Stud. 28: 178–185.
Nielsen, H. B., Sonne, A. M., Grunert, K. G., Banati, D., Pollak-Toth, A., Lakner, Z. (2009). Consumer per-
ception of the use of high-pressure processing and pulsed electric field technologies in food production.
Appetite. 52: 115–126.
Oey, I., Van der Plancken, I., Van Loey, A., Hendrickx, M. (2008). Does high pressure processing influence
nutritional aspects of plant based food systems? Trends Food Sci. Tech. 19: 300–308.
Olsen, N. V., Grunert, K. G., Sonne, A. M. (2010). Consumer acceptance of high-pressure processing and
pulsed-electric-field: A review. Trends Food Sci. Tech. 21: 464–472.
Onyango, B., Govindasamy, R., Hallman, W. (2006). US public awareness and knowledge of and interest in
biotechnology: A principal component factor analysis. J. Food Distr. Res. 37: 126–132.
Pecher, A., Tregear, A. (2001). Product country image effects for food products: The case of German cheese
in the UK. J. Int. Food Agribusiness. Marktng. 11(3): 1–15.
Pohlman, A. J., Wood, O. B., Mason, A. C. (1994). Influence of audiovisuals and food samples on consumer
acceptance of food irradiation. Food Tech. 48(12): 46–49.
Ravishankar, S., Zhang, H., Kempkes, M. L. (2008). Pulsed electric fields. Food Sci. Tech. Int. 14: 429–432.
Rimal, A. P., Moon, W., Balasubramanian, S. (2005). Agro-biotechnology and organic food purchase in the
United Kingdom. British Food J. 107: 84–97.
Rollin, F., Kennedy, J., Wills, J. (2011). Consumers and new food technologies. Trends in Food Sci. Tech. 22(2):
99–111.
Ronteltap, A., van Trijp, J. C. M., Renes, R. J., Frewer, L. J. (2007). Consumer acceptance of technology-based
food innovations: Lessons for the future of nutrigenomics. Appetite. 49: 1–17.
Rosenthal, A., Silva, J. L. (1997). Foods under pressure. Food Eng. 14: 37–39.
Sampedro, F., Rodrigo, M., Martinez, A., Rodrigo, D., Barbosa-Cánovas, G. (2005). Quality and safety aspects
of PEF application in milk and milk products. Critical Rev. Food Sci. Nutr. 45: 25–47.
Scholderer, J., Frewer, L. J. (2003). The biotechnology communication paradox: Experimental evidence and
the need for a new strategy. J. Cons. Policy. 26: 125–157.
Schutz, H. G., Bruhn, C. M., Diaz-Knauf, K. V. (1989). Consumer attitude toward irradiated foods: Effects of
labeling and benefit information. Food Tech. 43: 80–86.
Schutz, H. G., Cardello, A. V. (1997). Information effects on acceptance of irradiated foods in a military
population. Dairy Food Env. Sanit. 17(8): 470–481.
Selcuk, M., Oksuz, L., Basaran, P. (2008). Decontamination of grains and legumes infected with Aspergillus
spp. and Penicillium spp. by cold plasma treatment. Bior. Tech. 99: 5104–5109.
Sera, B., Gajdova, I., Cernak, M., Gavril, B., Hnatiuc, E., Kovacik, D., Kriha, V., Slama, J., Sery, M., Spatenka, P.
(2012). How various plasma sources may affect seed germination and growth. Optimization of Electrical
and Electronic Equipment, 13th International Conference on IEEE. 1365–1370.
Sharma, A., Collins, G., Pruden, A. (2009). Differential gene expression in Escherichia coli following expo-
sure to non-thermal atmospheric pressure plasma. J. App. Micr. 107: 1440–1449.
Siegrist, M., Keller, C., Kiers, H. A. L. (2006). Lay people’s perception of food hazards: Comparing aggre-
gated data and individual data. Appetite. 47: 324–332.
Sizer, C. E., Balasubramaniam, V. M., Ting, E. (2002). Validating high-pressure processes for low-acid foods.
Food Tech. 56: 36–42.
Sloan, A. E. (2003). What consumers want––and don’t want––on food and beverage labels. Food Tech. 57(11):
26–36.
Soliva-Fortuny, R., Balasa, A., Knorr, D., Martin-Belloso, O. (2009). Effects of pulsed electrical fields on
bioactive compounds in foods: A review. Trends Food Sci. Tech. 20: 544–556.
Sonne, A. M., Grunert, K. G., Olsen, N. V., Granli, B. S., Banati, D., Pollak-Toth, A. (2009). Consumer per-
ception of the use of high-pressure processing and pulsed electric field technologies in food production.
In: International European Forum on System Dynamics and Innovation in Food Networks, February
15–20, 2009, Austria (No. 59111). International European Forum.
Sonne, A. M., Grunert, K. G., Olsen, N. V., Granli, B. S., Szabó, E., Banati, D. (2012). Consumers’ perceptions
of HPP and PEF food products. British Food J. 114: 85–107.
Sorenson, D., Henchion, M. (2009). Consumers’ perceptions of novel process technologies: The case of
high pressure processed chilled ready meals. 113th European Association of Agricultural Economists
Seminar, A resilient European food industry and food chain in a challenging world 3rd September–6th
September 2009 Chania, Crete: Greece.
Sorenson, D., Henchion, M. (2011). Understanding consumers’ cognitive structures with regard to high pressure pro-
cessing: A means-end chain application to the chilled ready meals category. Food Quality Pref. 22: 271–280.
Sparks, P., Shepherd, R. (1994). Public perceptions of the potential hazards associated with food production
and food consumption: An empirical study. Risk Anal. 14: 799–806.
Swientek, B. (1999). New formulas for food science. Prepared Foods. 168: 33–40.
Tappi, S., Gozzi, G., Vannini, L., Berardinelli, A., Romani, S., Ragni, L., Rocculi, P. (2016). Cold plasma treat-
ment of fresh cut melon stabilization. Inn. Food Sci. Emerg. Tech. 33: 225–233.
Teisl, M. F., Fein, S. B., Levy, A. S. (2009). Information effects on consumer attitudes toward three food tech-
nologies: Organic production, biotechnology, and irradiation. Food Quality Pref. 20: 586–596.
Tuorila, H., Cardello, A. V., Lesher, L. (1994). Antecedents and consequences of expectations related to fat-
free and regular-fat foods. Appetite. 23: 247−263.
Vardag, T., Dierkes, H., Koener, P. (1995). High pressure for food processing. Food Tech. Euro. 2(3): 106–110.
Vega-Mercado, H., Martin-Belloso, O., Chang, F. J., Barbosa-Cánovas, G., Swanson, B. (1996). Inactivation of
Escherichia coli and Bacillus subtilis suspended in pea soup using pulsed electric fields. J. Food Proc.
Pres. 20(6): 501–510.
Vega-Mercado, H., Martin-Belloso, O., Qin, B. L., Chang, F. J., Gongora-Nieto, M. M., Barbosa-Canovas,
G. V. (1997). Non-thermal food preservation: Pulsed electric fields. Trends Food Sci. Tech. 8: 151–157.
Zimmerman, L., Kendall, P., Stone, M., Hoban, T. (1994). Consumer knowledge and concern about biotech-
nology and food safety. Food Tech. 48: 71–77.
Index
Note: Page numbers in italic and bold refer to figures and tables respectively.
Acidothermophilic bacteria 72 Canadian Food Inspection Agency (CFIA) 425

actin protein 56 Capsicum annuun (red bell pepper) 35
adiabatic heat of compression 375 carotenoids 245
advanced oxidation reduction process (AORP) 319 casein particles 75–6
agri-food wastes 30 cavitation bubbles 145, 146
alginate coating (ALC) 348 CCP (colloidal calcium phosphate) 75–6
Alicyclobacillus acidoterrestris 353 celiac disease 13, 15
alkaline phosphatase (ALP) 135–6 cell membrane 71
analytical ultrasound 160–2 cell rupture 238
Animal and Plant Health Inspection Service (APHIS) 435 cereal grains: eBeam 320–1; extraction, high pressure
animal products; see also meat and poultry products; effects on 31–2; HHP application on 12–13;
meat and poultry products, HPP impacts: PL HPP 13–14; industry 294–5
applications 183; quality 159–60 CFIA (Canadian Food Inspection Agency) 425
anisotropic susceptibility 266 CFMF (cross-flow microfiltration) 358
antimicrobials: EBP with 352; effect of ozone 190; HHP chain degradation 403
with hurdle effect combination 340–1; ozone cheese ripening process 81–2
with 351; PEF with 350; pulsed light processing chemical methods 190
with 347–8; ultrasonication with 343–4 chemical treatment: chlorine 227; hydrogen peroxide 228;
antioxidant compounds 419 NAI 228; organic acids 226–7, 227
AORP (advanced oxidation reduction process) 319 chemiclearance 425
APHIS (Animal and Plant Health Inspection Service) 435 Chilean papaya seeds, antioxidant capacity 30, 31
APPJ (atmospheric pressure plasma jet) 287, 300 chlorine, ozone/chemical treatment 227
aqueous ozone 225 circular dichroism (CD) analysis 114
areal density 317 civilization diseases 107
Arrhenius relationship 3 Clostridium botulinum 2–3, 380
ascorbate oxidase 419 Clostridium perfringens 343
aseptic food packaging 423 Clostridium sporogenes 337–8
Aspergillus flavus 344 codex of federal regulations (CFR) 425
Aspergillus niger 408 cold plasma technology 193, 444–5
Atlantic salmon 383 colloidal calcium phosphate (CCP) 75–6
atmospheric pressure discharge plasma 288 colostrum 82–3
atmospheric pressure plasma jet (APPJ) 287, 300 combination of preservation 331
autofrettage process 5 conventional pulse generation techniques 94–5
cool pasteurization (CP) 287, 378
Bacillus amylolique faciens 380 corona discharge 193, 215–16, 216
Bacillus cereus 353 covalent bonds 55, 75
Bacillus stearothermophilus 339 cross-flow microfiltration (CFMF) 358
Bacillus subtilis 337–8, 343 cross-linking 403
bacterial cells 71, 73
bakery applications, SCFX 248 dairy industry, HPP applications in 70, 80, 81; on cheese
barrier/silent discharge 288 making 80–2; dairy foods 82–3; water
barrier technology 331 molecules 75; yoghurt and ice-cream 82
batch operation mode 343 dairy products, PL applications 182
batch systems 148 Davis, A.R. 266
batch-type systems 70–1 DC glow discharge plasma 287–8
β-glucosidase (β-GLUC) 114 Defence Food Research Laboratory (DFRL), Mysore 121,
β–lactoglobulin 76–7, 136 122
bidirectional flyback converter 95, 96 dense phase carbon dioxide (DPCD) 236
biological factors, PEF 97 depressurization, effects 240
biological window effect 273–4 dielectric barrier discharge plasma 288
bipolar pulses 94, 110 dielectric breakdown theory 91, 91, 115
Botrytis cinerea 347 differential scanning calorimetry (DSC) 55–6, 56
bovine serum albumin (BSA) 77–8 DPCD (dense phase carbon dioxide) 236
455
456 inDeX
EAA (essential amino acid) 18 Food and Drug Administration (FDA) 436
eBeam see electron beam (eBeam) food-borne illnesses 417
EBP see electron beam processing (EBP) food borne pathogens 58–9
EDTA (ethylene diamine tetra acetic acid) 54, 340 food composition 178, 179, 180
egg and milk products, eBeam 321–2 Food Drug and Cosmetic Act 401
electrical breakdown 132 food irradiation 434–7, 435; global status 317; history 316
electrical discharge method 193 food matrix 408
electrical switch 109–10 food packaging industry 301–3, 303
electric field/energy, food preservation using 108 food preservation method 48
electric field strength (E) 96, 120 food processing operations 72
electrochemical method 193 food processing technologies, innovations 445–50, 448–9
electrochemical model 132 food products, irradiation 419
electrodes, configuration 92, 93 food quality, application on: food products 102–3; fruits
electron beam (eBeam) 434; advantages 315; bacterial and vegetable products 101–2; milk and milk
response 319; benefits 317; cereal products 320–1; products 98, 101
contemporary challenges 317; egg and milk food quality attributes, HPCD 250–1
products 321–2; FDA-approved applications 318; food safety 173, 190
fruits and vegetables 319–20; meat and poultry Food Safety and Inspection Service (FSIS) 435
products 321; moisture content 318; parameters FOS (fructo-oligosaccharides) 161
316–18; regulations and standards 323–5, 323, Fourier transform infrared spectroscopic (FTIR) 294
324; space food 322–3; spices 322 free fatty acids (FFA) 78
electron beam processing (EBP) 352; with antimicrobials 352 fructo-oligosaccharides (FOS) 161
electroporation theory 90–1, 91, 115 fruit juices, ozone application 199–200
electro-pulse technology 108 fruits and vegetable products, PEF 101–2; consumption 107;
“electropure” process 130 enzymes 113–15; microbial inactivation 111–13;
enzyme inactivation, HPCD: depressurization 240; rheological properties 117; textural characteristics
food ingredients 242; mechanisms 240; 115–17; toxin reduction effect on 119–21
microbubbled CO2 242; molecular CO2 239–40; fruits and vegetables: eBeam 319–20; PL applications 180,
pH 242; pH lowering effect 240; pressure 182; processing industry 298–9
cycles 242; pressure levels 241; processing fruits and vegetables, ozone application: microbial safety
media 242; temperature effect 241–2; treatment 196–7; nutrients, effect on 197–8; pesticide
time effect 242 residue 198–9; quality 197
enzymes 21, 382; activity 291; inactivation 114; vegetables FSIS (Food Safety and Inspection Service) 435
and fruits 113–15 FTIR (Fourier transform infrared spectroscopic) 294
Erwinia carotovora 444 functional and nutritional properties, PL applications 183–4
Escherichia coli 385, 420 Fusarium culmorum 408
essential amino acid (EAA) 18
essential oils (EOs) 341 gamma irradiation 320
“etching” process 290 gamma radiation 434
ethylene diamine tetra acetic acid (EDTA) 54, 340 gelation 159–60, 380
ethylene vinyl alcohol (EVOH) 400 generally recognized as safe (GRAS) 214, 350
exponential decaying pulses 94, 110 GH (growth hormone) 82
extraction, high pressure effects on 30; cereals 31–2, 32; ginsenosides yield 32, 33
fruits/vegetables and by-products 30; herbs glass packaging material 403
and roots 32; plant products 33; seafood and globular proteins 2–3
by-products 33 gluten, HHP effect on 15–16, 16
extrinsic factors 336; concentration 216–17; temperature 217 grain storage, ozone application: microorganisms 201;
extruded snacks, SCFX 249 mycotoxin 201; pests/insects 200–1; quality 202
gram-negative microorganisms/bacteria 71–2, 238, 340
fatty acids 54 gram-positive microorganisms/bacteria 71–2, 196, 238
FDA (Food and Drug Administration) 436 grape marc production 30
Fermi’s equation 114 GRAS (generally recognized as safe) 214, 350
FFA (free fatty acids) 78 growth hormone (GH) 82
fibrous proteins 3
fish, ozone application: microorganisms 204–5; quality hazard analysis and critical control point (HACCP) 321,
205–6 417, 445
flavanoids 245–6 heat coagulation time (HCT) 79
flavor compounds 246 Henry’s law 191
flexible packaging material 406 herb/spice processing industry 296–8
food additives 401 HHP see high hydrostatic pressure (HHP)
inDeX 457
HHP with hurdle effect combination: with acidification high-pressure sub-zero temperature (HPST) 36
(low pH) 337–8; with antimicrobials high-pressure treatment 70
340–1; with heat 338–40; with preservation high-temperature short-term (HTST) pasteurization 139
technologies 336–7 HILP (high-intensity light pulses) 355
hierarchical value maps (HVM) 438 HIPEF see high-intensity pulsed electric field (HIPEF)
high-dose treatment 318 homogeneous MFs 267
high-energy applications 435 horseradish peroxidase (HRP) 114
high hydrostatic pressure (HHP) 11, 418; advantage 12; HPCD see high-pressure carbon dioxide (HPCD)
aerobic mesophilic bacteria/molds and yeasts 22; HPF (high-pressure freezing) 36
on antinutritionals in legumes 19–20; application HPP see high-pressure processing (HPP)
29; and carbon dioxide 353–4; on cereals and HPST (high-pressure sub-zero temperature) 36
legumes 12–14; effectiveness 22; foodstuffs to HPU (high-power ultrasound) 341
13; and gamma irradiation 354; on gluten 15–16, HTST (high-temperature short-term) pasteurization 139
16; on grains 20–1; and MF 353; operation unit Hulsheger’s equation 114
49; and ozone 353; PEF and 358–9; phytic acid/ hurdle technology 373
phenolics, reduction in 20, 20; on protein and HVM (hierarchical value maps) 438
amino acids 17–18; raw soybean application 18; hydraulic fluid 376
from rice grains 13; on starch 14–15; TiO2-UV hydrogen bonds 55
photocatalysis 359–60; ultrasonication and 356; hydrogen peroxide 228
uses 12; on wheat dough 14 hydrophobic interactions 75
high-intensity light pulses (HILP) 355 hydrostatic pressure 381
high-intensity pulsed electric field (HIPEF) 108, 112–13, hydroxyl radical 290–1
131; application 118; effects 114–15
high osmotic pressure 354 ILP (intense light pulses) 374
high-power ultrasound (HPU) 341 immunoglobulins (Igs) 78, 82
high-pressure carbon dioxide (HPCD) 236; advantages implicit factors 336
238; application in beverage industry 242, 243, in-depth consumer research 450
244; application, water disinfection 251; cell industrial electron accelerators 434
rupture 238; critical point 236; disinfection of insulin-like growth factors-1 (IGF-1) 82
stored grains 251–2; displacement of oxygen intense light pulses (ILP) 374
237; enzyme inactivation 239–42; food intermediate-dose treatment 318
quality attributes 250–1; limitations 252–3; intrinsic factors 336; organic matter content 217; pH 217
microorganisms effects 238–9; nutraceuticals in-vitro protein digestibility 17
see nutraceuticals extraction; phase diagram ion cyclotron resonance model 268
237; regulatory aspects 253; SCFX 247–50, ionic bonds 75
247; solubilization and acidification 237–8 ionization radiation 402
high pressure effects: on dehydration 33–5; on extraction ionizing energy 424
30–3; on freezing 36; on frying 38; on infusion ionizing radiation 434
29, 29; on rehydration 35–6; on thawing 37–8 ion parametric resonance (IPR) 268
high-pressure extraction 30, 31 irradiation 401–3; packaging 403–6
high-pressure freezing (HPF) 36 isolation process 279
high-pressure pasteurization 396 isostaticity 375
high-pressure processing (HPP) 1, 375, 396–7, 437–9, isostatic pressing principle 2–3
440–2; advantages 376; application in food isostatic rule 28, 396
preservation 376; applications in meat, poultry,
and fish 48, 58–9; benefits and limitations 38–9; Langmuir, I. 285
cereals 13–14; on colour effect 383; combined Le Chatelier’s principle 2, 18, 28, 70, 375, 396
treatments with 59, 59–61; cycle 6–7; drying legumes protein: antinutritionals in 19–20; application
curve 34; on enzymes affecting quality effects of HHP on 12–13; HHP effect on 17–18;
382–3; features 1; on gelation effect 380–1; HPP 16
instrumentation and controls 5–6; legumes 16; lethal effect 177, 385
on lipid oxidation effect 382; manufacturing lipid oxidation 52–4, 382
units 4; meat tenderization by 53; on microbial lipoprotein lipase (LPL) 79
inactivation, effect 379–80; microfiltration lipoxygenase (LOX) 114
by 82–3; packaging requirement 397–401; liquid whistle 149
potential application 381; preservative effect 27; Listeria innocua 342, 348, 350, 385
pressure-creating device 5; pressure vessel 4–5; Listeria monocytogenes 341, 351, 354, 385, 444
pre-treatment 38; principles effects on food 2–3; low-dose treatment 318
technology 1–2; on texture effect 383–4; two end LOX (lipoxygenase) 114
closures 5; working principle 70–1; yoke 5 LPL (lipoprotein lipase) 79
458 inDeX
MA (malic acid) 348 microwave-driven remote plasma treatment 298

magnetic fields (MFs) 262; application 262; circular path microwave-powered CP treatment 297
265; definition 265; helical movement 265; milk and milk products, HPP: constituents 74, 74;
quality control 278–80; types 266–7 functional characteristics 79–80; on
magnetic freezing (MF) 275, 276–8 microorganisms 71–3; on milk fat, lactose, and
magnetic resonance imaging (MRI) 279–80 micronutrients 78; overview 69–70; on proteins
magnetic separation techniques 279 75–8; on water and minerals 75
magnetism 265 milk and milk products, PEF 98, 101, 131–2; applications
Maillard browning reactions 52–3 139–40; bacterial spores inactivation 133–4;
Maillard reaction products (MRP) 161–2 changes on milk constituents 136–7; enzymes
malic acid (MA) 348 134–6; physico-chemical parameters 137;
manosonication (MS) 356 sensory property 138; shelf-life 133
mano-thermo-sonication (MTS) 333, 357 milk enzymes, HPP-mediated effect on 78–9
MAP (modified atmospheric packaging) 407 milk fat globule (MFG) 137
meat and poultry products: eBeam 321; processing milk pasteurization 80
industry 300–1 milk processing industry 299–300
meat and poultry products, HPP impacts 49–50; changes mixed dimer formation 178
50; on colour 50–2; effects 49; on flavour 52–3; modified atmospheric packaging (MAP) 407
on lipids 53–4; microflora in 58; on non-heme molecular CO2 239–40
iron 54; principle and mechanism 48–9; on molecular ozone 215
proteins 54–5; on texture 53; on thermal Monilia fructigena 347
behaviour 55–6; on WHC 56–7 Moses effect 275
meat, ozone application: microorganisms 202–3; quality MRI (magnetic resonance imaging) 279–80
203–4 MRP (Maillard reaction products) 161–2
media factors, PEF 97–8 MS (manosonication) 356
medium-energy applications 435 MTS (mano-thermo-sonication) 333, 357
MFs, food preservation: isolation and separation 279; multifactorial preservation 331
magnetic freezing 275, 276–8; microbial multi-targeted approach 332
inactivation 268–9, 271–2; microorganisms mussel meat 379
and enzymes 267–8, 270–1; MRI 279–80; mycotoxins 120; effects 201; in stored grains 200
pasteurization 272; sterilization 272–5, 274; yeast myosin 380–1, 383
under high-gradient magnetic field 275, 279
microbial cells 71–2 nanoclusters/nanotubes 76
microbial destruction 50 native myosin 380
microbial inactivation 21–3, 73, 97, 385; HPCD 238–9; negative air ions (NAI) 228
kinetic models for PEF 99–100; mechanism net effects 3, 336
90–1, 91; MFs 268–9, 271–2; OMF 269, 271–2; Newtonian behaviour 117
ozone 194, 195; pH influence on 98; plasma 288, nisin, bacteriocins 340, 350
289–90, 290–1; PMF 268; principles 130–1; NMR (nuclear magnetic resonance) 36, 267
SMF 269, 271; sonication 150; synergistic effect non-Newtonian fluids 117
150–1; thermosonication 150–1; ultrasonication non-thermal food processing 262, 263–4
152–3; vegetables and fruits 111–13 non-thermal plasma (NTP) 287–8, 289–90, 444;
microbial inactivation by PL: distance from light source functional modification of food 292
180; food composition 178, 179, 180; food non-thermal preservation techniques 374–5
industry, applications 180, 181–2, 182–4; non-thermal processed foods: commercial applications
microorganism type 178 426–8; consumer response 423–4; energy
microbial quality of juices 220–2 requirements/environmental concerns 420–2;
microbial safety, effect 196–7 nutritional aspects 418–20; overview 417–18;
microbubbled CO2 242 packaging issues 422–3; regulatory aspects 424–6
microfiltration (MF) 82–3; HHP 353; PEF 358 non-thermal processes and hurdle effects: concept 331–4,
microflora reduction 224 332, 333, 334; with conventional hurdles
microorganisms 22–3, 97; and enzymes, effects 267–8, 336–52; future scope 361–2; hurdle approach in
270–1; fish 204–5; grains storage 201; gram- 352–60; overview 330–1
positive 71–2; HPCD effects 238–9; HPP effect non-thermal processing/processes 418, 422, 443
on 57, 57; HPP on milk 71–3; inactivation 109; NTP see non-thermal plasma (NTP)
and initial load 196; meat 202–3; microbial nuclear magnetic resonance (NMR) 36, 267
inactivation 238–9; pathogenic 73, 375; nutraceuticals extraction: carotenoids 245; flavanoids
probiotic 83; resistance 71; types 178 245–6; pressure, effect 245; SC-CO2 247;
microscopic ordering principle 3 temperature, effect 244
microwave discharge plasma 288 nutrients, effect on 197–8
inDeX 459
ohmic heating 130, 330 physico-chemical properties, milk 137

oligomer 380 phytosanitary irradiation 316
olive pomace production 30 piezophiles 2
organic acids 226–7, 227 PL see pulsed light (PL)
oscillating/pulse magnetic field (OMF) 266–7, 269 PL applications, food industry 181–2; animal products 183;
oscilloscope 109 dairy products 182; demerits 184; fruits and
osmotic/osmo-dehydration process 35, 101 vegetables 180, 182; functional and nutritional
oxygen, displacement 237 properties 183–4; merits 184; sea food 183
oysters 378–9 plasma application: in cereal industry 294–5; food
ozone 419; antimicrobial effect 190; with antimicrobials packaging industry 301–3, 303; food processing
351; application in food commodities 196–206; industry 299; fruits and vegetables processing
with conventional preservation technologies industry 298–9; herb/spice processing industry
350–1; extrinsic factors 216–17; factors 296–8; meat and poultry processing industry
affecting 194–6; generation 193–4; GRAS 300–1; milk processing industry 299–300; seed
190; with heat 351; HHP and 353; intrinsic germination 296
factors 217; molecular structure 191; PEF plasma efficiency, factors affecting 303–4: direct and
358; physico-chemical characteristics 191–3, indirect treatment 304; gas/mixture of gases
192; as preservative 218–19, 219; safety and 304; microbial inactivation 305
risks 206; structure 191; synthesis 191; and plasma in food processing 444; application see plasma
ultrasonication 360 application; classification 286–8, 286;
ozone application: constraints 228; fish 204–6; in food constituents 285; definition 284; disinfection
processing 214; fruit juices 199–200; fruits and 284; efficiency affecting factors 303–5; enzyme
vegetables 196–9; grain storage 200–2; GRAS activity 291, 292; in food processing see plasma
214; meat 202–4 in food processing; functional properties 293;
ozone, factors affecting: microorganisms and initial ionization 286; limitations 305–6; microbial
load 196; organic components 196; pH 195; inactivation 288, 289–90, 290–1; net neutral
temperature 194 charge 285; seed germination 292–3; stages of
ozone generation: corona discharge 215–16, 216; matter 285; starch modification 291–2
electrical discharge 193; electrochemical 193; plastic films 399
microbial inactivation mechanism 194, 195; PL processing: with antimicrobials 347–8; with
photochemical 216; radiation 193–4 conventional preservation hurdles 346; with
heat 347; ultrasonication and 355–6
packaging requirement: for HPP 397–400; for irradiation PLT (pulsed light technology) 344, 385
403–6, 404, 405; for PEF 397–8, 407; for PL PME (pectin methylesterase) 113, 139, 154
treatment 408–9 PMF see pulsed magnetic field (PMF)
Panax ginseng 32 POD (peroxidase) 113, 153
parallel plate systems 92 polyethylene terephthalate (PET) 400
Pascal’s isostatic principle 2, 375 polymer foaming 250
pasteurization 272, 376, 395 polymeric materials 301–2, 303
pathogenic microorganisms 73, 375 polyphenol oxidase (PPO) 153, 385
PATP (pressure-assisted thermal process) 2–3, 378, 384 polyunsaturated fatty acids (PUFA) 382
PATS (pressure-assisted thermal sterilization) 333, 374 polyvinyl alcohol (PVOH) 400
PDA (potato dextrose agar) 120–1 polyvinyl chloride (PVC) 408
pectinesterase (PE) enzymes 114 “PoroCrit” process 245–6
pectin methylesterase (PME) 113, 139, 154 potato dextrose agar (PDA) 120–1
PEF see pulsed electric field (PEF) power electronics techniques 95–6
PER (protein efficiency ratio) 58 power ultrasound 147, 160, 162
peroxidase (POD) 113, 153 PPO (polyphenol oxidase) 153, 385
pesticide residues: effect on 120; fruits and vegetables preservation, ozone 218–19, 219; chemical treatment
198–9; grain storage 200–1 226–8, 227; combination treatments 219,
PET (polyethylene terephthalate) 400 220–2; physical treatment 223–5, 223,
PFN (pulse forming networks) 92, 94 224, 226
phase transition 396 pressure-assisted thermal process (PATP) 2–3,
pH lowering effect 240 378, 384
photochemical ozone generator 216 pressure-assisted thermal sterilization (PATS) 333, 374
physical methods 190 pressure cycles 242
physical treatment: PEF 225; thermal treatment 225, 226; pressure levels 241
ultrasound 223–4, 223; UV radiation 224–5, 224 pressure shift freezing (PSF) 36, 37
physico-chemical characteristics, ozone 192; solubility pressure treatment, mechanism 375–6
191–2, 192; stability 192–3 pressurisation/pressurization 51, 75, 384
460 inDeX
probiotic microorganism 83 quality effects on: fishes 205–6; fruits and vegetables 197;
processing factors 336 grain storage 202; meat 203–4
process variables 407
prokaryotes 337 radiation method 193–4
protein denaturation 18, 19, 55; enzymes 21; HHP on radio frequency (RF) 287; discharge plasma 288
antinutritionals in legumes 19–20; HHP on Raman spectroscopy 50–1
grain 20–1; microbial inactivation 21–3; raw chevon muscle, colour changes in 51, 52
sarcoplasmic 55 Rawls, J. R. 266
protein efficiency ratio (PER) 58 raw milk cheese 138
proteins: food 75; HPP impacts on 54–5; HPP on milk RCT (rennet coagulation time) 79–80
75–8; versus myofibrillar proteins 56 reactive oxygen species (ROS) 359
proteolysis process 77, 79 red bell pepper (Capsicum annuun) 35
proteolytic enzymes 383 regulations and standards, eBeam: European Union
Pseudomonas aeruginosa 269 countries 324; Indian regulations, allied
Pseudomonas bacteria 72, 101 product 324; Indian regulations, food classes
PSF (pressure shift freezing) 36, 37 323; Radura symbol 324, 324
PUFA (polyunsaturated fatty acids) 382 regulatory aspects, HPCD 253
pulsed discharge plasma 288 rehydration 155; high pressure effects on 35–6
pulsed electric field (PEF) 225, 406–7, 440–2, relative residual activity (RRA) 135
443–4; with acidic pH 349–50; on aflatoxin rennet coagulation time (RCT) 79–80
production 112; with antimicrobial 350; response surface methodology (RSM) 115
application 102; bacterial spores inactivation RF see radio frequency (RF)
in milk 133–4; components 131; with rheological analysis 117
conventional preservation hurdles 348–9; ROS (reactive oxygen species) 359
dairy products applications 139–40; RRA (relative residual activity) 135
definition 130; at DFRL, Mysore 121; RSM (response surface methodology) 115
equipment 91; factors to effectiveness 96–8;
with heat 349; and HHP 358–9; and HPCD SAA (supercritical-assisted atomization) process 247
357–8; inactivation 97; kinetic models for 98; Saccharomyces cerevisiae 338, 343, 350
and MF 358; microbial inactivation principles Salmonella enterica typhimurium 357
130–1; milk and milk products 131–2; Salmonella enteritidis 385
milk constituents 136–7; milk enzymes sarcoplasmic protein fraction 55
134–6; overview 89–90; and ozone 358; SASP (small acid-soluble proteins) 347
packaging requirement 407; parameters Saudavel com maissabor 439
110–11, 110; physico-chemical and sensory scanning electron microscopy 14
properties 137–8; as pre-treatment aid 119; SC-CO2/SCCD see supercritical carbon dioxide
principle 90; processing 108–10; pulse (SC-CO2/SCCD)
generation networks 94–6; pulses types SCFE (supercritical fluid extraction) 244
and characteristics 92–4, 93; set up 92; SCFX see supercritical fluid extrusion (SCFX)
shelf stability 118; synergistic effect 138–9; seafoods: fish 376–8; PL applications 183; pulsed light in
treatment chamber 92, 109; treatments 97–8, 385–6
101–3, 111, 116, 116; unit operations layout sealing parameters 401
109; vegetative microbial cells 132–3 seed germination 292–3, 296
pulsed light (PL) 407–8; generation 175–6, 176; sensory property, milk 138
mechanism of action 177–8, 178; merits shelf life extension 48, 378–9
and demerits 184; microbial inactivation shellfish 378–9
see microbial inactivation by PL; packaging small acid-soluble proteins (SASP) 347
requirement 408–9; principle 174–5, 175; SMFs (static magnetic fields) 267, 269
UV-C 174 soft laddering system technique 437
pulsed light technology (PLT) 344, 385 solubilization and acidification 237–8
pulsed magnetic field (PMF) 262; in food preservation space food, eBeam 322–3
see MFs, food preservation; types 266–7 spices, eBeam 322
pulsed UV disintegration 409 spontaneous reactions 70
pulse forming networks (PFN) 92, 94 spores inactivation 133–4
pulse generation networks: bipolar pulses 94; conventional square wave pulses 132
techniques 94–5; exponential decaying pulses stages of matter 285
94; oscillatory pulses 94; power electronics Staphylococcus aureus 342
based technique 95–6 starch, HHP effect on 14–15
PVC (polyvinyl chloride) 408 starch modification 291–2
PVOH (polyvinyl alcohol) 400 static magnetic fields (SMFs) 267, 269
inDeX 461
sterilization: biological window effect 273–4; ultrasonic-assisted extraction of biomolecules

electromagnetic field 274; low-pressure PMF 155, 156–7
273–4, 274; technique 408 ultrasonic-assisted freezing (UAF) 158–9
stored grains, disinfection of 251–2 ultrasonication: with antimicrobials 343–4; with
superconducting technology 275 conventional preservation technologies 341–2;
supercritical-assisted atomization (SAA) process 247 with heat (thermo-sonication) 342–3; heat
supercritical carbon dioxide (SC-CO2/SCCD) 353–4; and HHP (MTS) 357; and HHP (MS) 356;
encapsulation using 247 and ionizing radiation processing 356; and
supercritical fluid extraction (SCFE) 244 osmotic pressure (osmo-sonication) 354–5;
supercritical fluid extrusion (SCFX) 247; advantages and PEF processing 356; and pulsed light
248; bakery 248; expansion mechanism 248; processing 355–6; and supercritical carbon
extruded snacks 249; polymer foaming 250; dioxide (SC-CO2) 354; and UV radiation
texturization 249–50 (photosonication) 355
surface discharge plasma 288 ultrasonic cavitation 148, 386
synergistic bactericidal effects 333 ultrasonics applications: animal product quality 159–60;
synergistic effect 139 batch-type probe 149; biomolecules, extraction
synthesis of ozone 191 155, 156–7; continuous probe 149; drying 155,
158; enzyme inactivation 153–4, 154; food
technological factors, PEF: duration of treatment 97; industry 160–2; microbial inactivation 150–2,
electric field strength (E) 96 152–3; UAF 158–9; ultrasonic bath 149
TEM (transmission electron microscopy) 76, 111 ultrasonics in food processing: applications 146; see
temperature: affecting enzyme inhibition 241–2; on ozone also ultrasonics applications; bath and probe
efficacy 194 system 148; cavitation bubbles 145, 146;
texturization 249–50 engineering aspects 147–9; equipment 148;
thermal-based food processes 421 frequency ranges 147; principles 147–9;
thermal plasma (TP) 286–7, 444 rarefaction 145
thermal processing 395 ultrasonic transducers 147
thermal treatments 129–30, 225, 226, 417; shelf life 262 ultrasound 223–4, 223, 355; analytical 160–1; applications
thermo-sonication (TS) 342 160, 162; effects 160; in food industry 147;
thymine-cytosine dimer 178 low-intensity 147; processing 386; to strawberry
thymine dimer 178 juice 150; treatment 151; uses 155
TMA (trimethylamine) content 205 ultrasound-assisted cutting 161
TMAO (trimethylamine oxide) 377 ultrasound-assisted mixing 161–2
total plate count (TPC) 379 ultrasound-assisted osmotic dehydration (UOD) 158
total volatile bases (TBA) 205 ultraviolet light assisted titanium dioxide photocatalysis
toxic microbial metabolites, effect on 120–1 (tiO2-UV photocatalysis) 359–60
toxin reduction in vegetable/fruits 119; pesticide residues ultraviolet (UV) radiation 224–5, 224; ozone generation
120; on toxic microbial metabolites 120–1 193–4; processing with hurdles 344, 345;
Trachurustrachurus 382 ultrasonication and 335
traditional pasteurization (TP) 378 UOD (ultrasound-assisted osmotic dehydration) 158
traditional thermal technology 395 UV radiation see ultraviolet (UV) radiation
transforming growth factor beta-2 (TGF-β2) 82
transition state theory (TST) 70 vegetative microbial cells 132–3
transmission electron microscopy (TEM) 76, 111 viruses 380
treatment chamber 70, 92
treatment gap 131 water disinfection, HPCD 251
trimethylamine (TMA) content 205 water holding capacity (WHC) 36; HPP impact on 56–7
trimethylamine oxide (TMAO) 377 water immersion thawing (WIT) 38
TS (thermo-sonication) 342 whey protein concentrate (WPC) 80
TST (transition state theory) 70 whey protein isolates (WPI) 137
UAF (ultrasonic-assisted freezing) 158–9 xanthohumol 32

ultra-high magnetic fields 267 X-ray irradiation technologies 434
ultra-high pressure (UHP) 1, 70
ultrasonic-assisted drying 155, 158 Zimmerman theory 132
ultrasonic-assisted enzyme inactivation 153–4, 154 Zygosaccharomyces bailii 338

Chauhan, O. P - Non-Thermal Processing of Foods-Chapman and Hall - CRC (2019)

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Chauhan, O. P - Non-Thermal Processing of Foods-Chapman and Hall - CRC (2019)

Uploaded by

Copyright:

Available Formats

Non-thermal Processing of Foods

© 2019 by Taylor & Francis Group, LLC

No claim to original U.S. Government works

Printed on acid-free paper

International Standard Book Number-13: 978-1-1380-3584-3 (Hardback)

Library of Congress Cataloging‑in‑Publication Data

Names: Chauhan, O. P., author.

Visit the Taylor & Francis Web site at

and the CRC Press Web site at

Bharat Bhushan S. J. Kale

R. Kumar Prerna Nath

Feby Luckose A. K. Pandey

Shruti Sethi Khudsia Sultana

Simran Kaur Arora and O. P. Chauhan

1.1 HPP Technology ....................................................................................................................... 1

1.1 hpp tEChNOLOGY

HPP is also known as High Hydrostatic Pressure Processing, Ultrahigh-Pressure Processing,

1.1.1 principles Governing the Effects of hpp on Food

1.1.2 Global Manufacturers of hpp Units

1. USA: Avure Technologies, Engineered Pressure Systems Incorporated, Harwood Engineering,

1.2 hIGh prESSUrE EQUIpMENt aND ENGINEErING aSpECtS

The HPP equipment typically consists of:

high hydrostatic pressure processing

Sajad Ahmad Wani and Pradyuman Kumar

2.1 Introduction ............................................................................................................................ 11

2.2 appLICatION OF hhp ON CErEaLS aND LEGUMES

2.2.1 high-pressure processing of Cereals

2.2.2 Effect of hhp on Starch

2.2.3 Effect of hhp on Gluten

2.2.4 high-pressure processing of Legumes

2.2.5 hhp Effect on Legume proteins

parameters raw Soybean hhp treated Soybean

Essential amino acids

table 2.1 (Continued) Effect of hhp on the total protein and

parameters raw Soybean hhp treated Soybean

Source: Penas, e. et al., Food Chem., 125, 423–429, 2011.

2.3 DENatUratION OF prOtEINS

2.3.1 Effect of hhp on the antinutritionals present in Legumes

pressure (Mpa), trypsin Inhibitor

Source: Linsberger-Martin, G. et al., LWT-Food Sci. Technol., 51, 331–336, 2013.

2.3.2 Influence of hhp on the Constituents of Grains

2.3.4 Microbial Inactivation

2.4 FUtUrE aSpECtS

Effect of high-pressure processing on

Jincy M. George and Navin Kumar Rastogi

3.1 Introduction ........................................................................................................................... 27

3.2 EFFECt OF hIGh prESSUrE ON INFUSION

3.3 EFFECt OF hIGh prESSUrE ON EXtraCtION

3.3.1 Fruits, Vegetables, and their By-products

DPPH (μmol TE g-1 seeds)

γ-Oryzanol contents (mg/100g, dry basis)

3.3.3 herbs and roots

Yield of ginsenosides (%)

3.3.4 Seafood and By-products

3.3.5 Other plant products

3.4 EFFECt OF hIGh prESSUrE ON DEhYDratION

1.2 Control 600 MPa/ 5 min 600 MPa/ 30 min

Control 600 MPa/ 5 min 600 MPa/ 30 min

3.5 EFFECt OF hIGh prESSUrE ON rEhYDratION

3.6 EFFECt OF hIGh prESSUrE ON FrEEZING

3.7 EFFECt OF hIGh prESSUrE ON thaWING

3.8 EFFECt OF hIGh prESSUrE ON FrYING