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Nanotechnology Applications
in the Food Industry
http://taylorandfrancis.com
Nanotechnology Applications
in the Food Industry

Edited by
V Ravishankar Rai
Jamuna A. Bai
CRC Press
Taylor & Francis Group
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Boca Raton, FL 33487-2742

© 2018 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: 978-1-4987-8483-2 (Hardback)

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 Contents
Preface .............................................................................................................................................. ix
Editors .............................................................................................................................................. xi
Contributors.................................................................................................................................... xiii

Part I
Introduction to Nanotechnology in the Food Sector

Chapter 1
Development of Bio-Based Nanostructured Systems by Electrohydrodynamic Processes...............3
Maria Jose Costa, Philippe Emmanuel Ramos, Pablo Fuciños, José António Teixeira,
Lorenzo Miguel Pastrana, and Miguel Ângelo Cerqueira

Chapter 2
Carbon Nanomaterial-Based Fertilizers Can Improve Plant Growth..............................................21
Guixue Song, Madelyn Pandorf, Paul Westerhoff, and Yun Ma

Chapter 3
Nanomaterials in Food Applications................................................................................................45
Ahmed S. Khan

Chapter 4
Market Potential of Food Nanotechnology Innovations..................................................................59
Rajesh P. Shastry and V Ravishankar Rai

Part II
Nanotechnology in Food Packaging

Chapter 5
Nanomaterials Applicable in Food Protection.................................................................................75
Josef Jampílek and Katarína Kráľová

Chapter 6
Use of Nanopolymers, Nanocomposites, and Nanostructured Coatings in Food Packaging..........97
Semih Ötleş and Buket Yalçın Şahyar

Chapter 7
Starch Nanocomposite Films for Food Packaging.........................................................................107
Oswaldo Ochoa Yepes, Lucas Guz, Santiago Estevez Areco, Roberto Candal,
Silvia Goyanes, and Lucía Famá

Chapter 8
Electrospun Nanofibers: Development and Potential in Food Packaging Applications................141
Carlos A. Fuenmayor and Paula J. P. Espitia

v
VI CONTENTS

Part III
Nanosensors for Safe and Quality Foods

Chapter 9
Nanosensors in the Food Industry.................................................................................................183
Fabrizio Sarghini and Francesco Marra

Chapter 10
Applications of Aptamers to Nanobiosensors and Smart Packaging............................................203
Luis Eduardo Suárez-Nájera, Stefany Cárdenas-Pérez, José Jorge Chanona-Pérez,
Arturo Manzo-Robledo, Jaime Vargas-Cruz, Mayra Luna-Trujillo,
Miriam Rodríguez-Esquivel, and Mauricio Salcedo-Vargas

Part IV
Nanotechnology for Nutrient Delivery in Foods

Chapter 11
Nanoemulsions Produced by Low-Energy Methods: Fundamentals and Food Applications.......221
Samantha C. Pinho, Cynthia de Carli, and Marilia Moraes-Lovison

Chapter 12
Application of Nanotechnology in the Safe Delivery of Bioactive Compounds..........................237
Behrouz Ghorani, Sara Naji-Tabasi, Aram Bostan, and Bahareh Emadzadeh

Chapter 13
Electrospinning of Edible, Food-Based Polymers.........................................................................293
Serife Akkurt, Lin Shu Liu, and Peggy Tomasula

Chapter 14
Role of Polymeric Nanoparticles in Nutraceutical Delivery..........................................................315
Rubiana Mara Mainardes and Najeh Maissar Khalil

Chapter 15
Nanoemulsions and Nanodispersions: A Fundamental View of Their Preparation,
Characterization, Stability Evaluation, and Application................................................................333
Tai Boon Tan and Chin Ping Tan

Chapter 16
Lipid Nanocarriers for Phytochemical Delivery in Foods.............................................................357
Bojana D. Balanč, Kata T. Trifković, Radoslava N. Pravilović, Verica B. Đorđević,
Steva M. Lević, Branko M. Bugarski, and Viktor A. Nedović
CONTENTS VII

Chapter 17
Role of Bioactive Compounds to Improve Nanomechanical Properties and Functionality
of Polymeric Vehicles....................................................................................................................385
Alonso Villafán-Rangel, Juan Eduardo René Cortés-Millán, Miriam Madrid-Mendoza,
Israel Arzate-Vázquez, Monserrat Escamilla-García, Ollin Celeste Martínez-Ramírez,
and Angélica Gabriel Mendoza-Madrigal

Chapter 18
Microscopy Techniques for Structural Characterization, Evaluation of Nanomechanical
Properties, and Biosensing of Food Systems.................................................................................403
Luis Eduardo Suárez-Nájera, Stefany Cárdenas-Pérez, Juan Vicente Méndez-Méndez,
José Jorge Chanona-Pérez, Alejandra Valdivia-Flores, Georgina Calderón-Domínguez,
and Rubén López-Santiago

Chapter 19
Chitosan Micro- and Nanoparticles for Vitamin Encapsulation....................................................427
Patricia Rosales-Martínez, Maribel Cornejo-Mazón, Izlia J. Arroyo-Maya,
and Humberto Hernández-Sánchez

Chapter 20
Nanoparticle Nutrient Carriers.......................................................................................................441
Nily Dan

Chapter 21
Electrospinning and Electrospraying Technologies and Their Potential Applications
in the Food Industry.......................................................................................................................461
Alex López-Córdoba, Clara Duca, Jonathan Cimadoro, and Silvia Goyanes

Part V
Safety Assessment for Use of Nanomaterials in Food and Food Production

Chapter 22
Advanced Approaches for Efficacy Evaluation and Risk Assessment of Nanomaterials in Food...483
Marco Roman, Catia Contado, Christian Micheletti, Iolanda Olivato, Erik Tedesco,
and Federico Benetti

Chapter 23
Effect of Nanoparticles on Gastrointestinal Tract..........................................................................503
Jamuna A. Bai and V Ravishankar Rai

Chapter 24
Regulatory Framework for Food Nanotechnology........................................................................517
Kangkana Banerjee and V Ravishankar Rai

Index.............................................................................................................................................. 529
http://taylorandfrancis.com
 Preface
Nanotechnology Applications in the Food Industry is a comprehensive reference book containing
exhaustive information on nanotechnology and the scope of its applications in the food industry. The
various sectors in the food industry where nanotechnology finds extensive application include the
production, processing, packaging, and preservation of foods. Nanotechnology is also increasingly
used to enhance flavor and color and in nutrient delivery and bioavailability and to improve food
safety and in quality management.
The book focuses on the overview of nanotechnology development for food industries and the
impact it is having on the state of science in the food industry. The book has five sections delving into
all aspects and the key role of nanotechnology in the food industry in the present scenario and the
future trends. Part I (Introduction to Nanotechnology in the Food Sector) covers the technological
basis for its application in the food industry and in agriculture, the use of nanosized foods and
nanomaterials in food, the safety issues pertaining to its applications in the foods, and on market
analysis and consumer perception of food nanotechnology. Part II (Nanotechnology in Food Pack-
aging) reviews the use of nanopolymers, nanocomposites, and nanostructured coatings in food
packaging. The potential of nanotechnology in active packaging applications, intelligent packaging,
and multifunctional packaging for improving the safety and quality of foods are discussed. Part III
(Nanosensors for Safe and Quality Foods) provides an overview on nanotechnology in the develop-
ment of biosensors for pathogen and food contaminant detection and in sampling and food quality
management. The challenges and trends in nanosensor design for food applications are discussed. Part
IV (Nanotechnology for Nutrient Delivery in Foods) deals with the use of nanotechnology in foods for
controlled and effective release of nutrients. The use of nanoparticles in developing, increasing the
functionality and bioactivity of dietary foods, functional foods, and nutraceuticals are covered. Part V
(Safety Assessment for Use of Nanomaterials in Food and Food Production) deliberates on the benefits
and risks associated with the extensive and long-term application of nanotechnology in the food sector.
The issues on nanoparticles and their residues in food during food production, migration of
nanoparticles into foods during processing and packaging, behavior of nanoparticles in the gastro-
intestinal tract, and safety and risk assessment of nanomaterials in the food are reviewed. Legal and
ethical barriers for the application of nanotechnology in food industry, regulatory aspects of its
application in food, and food production and labeling of nano ingredients in foods are addressed.
The book is a valuable reference material discussing the basics and trends in the application of
nanotechnology in the food industry. It is exhaustive and comprehensive, focusing on all aspects of
technological strategies in the nanofood segment. The book is beneficial for graduate students,
researchers, scientists working on application aspects of nanotechnology and food industry and food
policy makers.
We would like to thank all the authors for contributing the chapters and sharing their expertise.

ix
http://taylorandfrancis.com
 Editors
V Ravishankar Rai earned his MSc and PhD from the University of Mysore, India. Currently,
Dr. Rai is working as a professor in the Department of Studies in Microbiology, University of
Mysore, India. He was awarded a fellowship from the UNESCO Biotechnology Action Council,
Paris (1996), the Indo-Israel Cultural Exchange Fellowship (1998), the Biotechnology Overseas
Fellowship, Government of India (2008), the Indo-Hungarian Exchange Fellowship (2011), Indian
National Academy Fellowship (2015), and Cardiff Incoming Visiting Fellowship (2017). Presently,
he is the coordinator for the Department of Science and Technology, Promotion of University
Research and Scientific Excellence and University Grants Commission innovative programs.

Jamuna A. Bai has completed her MSc and PhD in microbiology from the University of Mysore,
India. She is working as a researcher in the University Grants Commission-sponsored University
with Potential Excellence Project, University of Mysore, India. She has previously worked as Indian
Council of Medical Research Senior Research Fellow and carried research on food safety, role of
quorum sensing and biofilms in food-related bacteria, and developing quorum-sensing inhibitors.
Her research interests also include antimicrobial application of functionalized nanomaterials against
food-borne pathogens.

xi
http://taylorandfrancis.com
 Contributors
Serife Akkurt Bojana D. Balanč
Dairy and Functional Foods Research Unit Faculty of Technology and Metallurgy
Eastern Regional Research Center Department of Chemical Engineering
Agricultural Research Service University of Belgrade
United States Department of Agriculture Belgrade, Serbia
Wyndmoor, Pennsylvania
Kangkana Banerjee
and Department of Studies in Microbiology
Rutgers University of Mysore
Department of Food Science Mysore, India
The State University of New Jersey
New Brunswick, New Jersey Federico Benetti
European Center for the Sustainable Impact
of Nanotechnology
Santiago Estevez Areco
EcamRicert Srl
Facultad de Ciencias Exactas y Naturales
Rovigo, Italy
Departamento de Física, Laboratorio
de Polímeros y Materiales Compuestos
Aram Bostan
(LP&MC)
Department of Food Nanotechnology
Instituto de Física de Buenos Aires
Research Institute of Food Science
(IFIBA-CONICET)
and Technology
Universidad de Buenos Aires
Mashhad, Iran
Buenos Aires, Argentina
Branko M. Bugarski
Izlia J. Arroyo-Maya Faculty of Technology and Metallurgy
Departamento de Procesos y Tecnología, Department of Chemical Engineering
Unidad Cuajimalpa University of Belgrade
Universidad Autónoma Metropolitana Belgrade, Serbia
Mexico City, Mexico
Georgina Calderón-Domínguez
Israel Arzate-Vázquez Departamento de Ingeniería Bioquímica
Laboratorio de Microscopía de Fuerza Atómica Escuela Nacional de Ciencias Biológicas
y Nanoindentación, Centro de Nanociencias Instituto Politécnico Nacional
y Micro y Nanotecnologías Mexico City, Mexico
Instituto Politécnico Nacional
Unidad Profesional Adolfo López Mateos Roberto Candal
Mexico City, Mexico Instituto de Investigación e Ingeniería
Ambiental, Consejo Nacional
Jamuna A. Bai de Investigaciones Científicas y Técnicas
Department of Studies in Microbiology Universidad Nacional de San Martín
University of Mysore Buenos Aires, Argentina
Mysore, India

xiii
XIV CONTRIBUTORS

Stefany Cárdenas-Pérez Nily Dan

Departamento de Ingeniería Bioquímica Department of Chemical and Biological
Escuela Nacional de Ciencias Biológicas Engineering
Instituto Politécnico Nacional Drexel University
Mexico City, Mexico Philadelphia, Pennsylvania

Miguel Ângelo Cerqueira Cynthia de Carli

International Iberian Nanotechnology Department of Food Engineering
Laboratory School of Animal Science and Food Engineering
Braga, Portugal University of Sao Paulo
Pirassununga, Brazil
José Jorge Chanona-Pérez
Departamento de Ingeniería Bioquímica Verica B. Đorđević
Escuela Nacional de Ciencias Biológicas Faculty of Technology and Metallurgy
Instituto Politécnico Nacional Department of Chemical Engineering
Mexico City, Mexico University of Belgrade
Belgrade, Serbia
Jonathan Cimadoro
Facultad de Ciencias Exactas y Naturales Clara Duca
Departamento de Física, Laboratorio Facultad de Ciencias Exactas y Naturales
de Polímeros y Materiales Compuestos Departamento de Física, Laboratorio de
(LP&MC) Polímeros y Materiales Compuestos (LP&MC)
Instituto de Física de Buenos Aires Instituto de Física de Buenos Aires
(IFIBA-CONICET) (IFIBA-CONICET)
Universidad de Buenos Aires Universidad de Buenos Aires
Buenos Aires, Argentina Buenos Aires, Argentina

Bahareh Emadzadeh
Catia Contado
Department of Food Nanotechnology
Department of Chemical
Research Institute of Food Science
and Pharmaceutical Sciences
and Technology
University of Ferrara
Mashhad, Iran
Ferrara, Italy
Monserrat Escamilla-García
Maribel Cornejo-Mazón Facultad de Química
Departamento de Biofísica, Escuela Nacional Universidad Autónoma de Querétaro
de Ciencias Biológicas del Instituto Santiago de Querétaro, Mexico
Politécnico Nacional
Unidad Profesional Lázaro Cárdenas Paula J. P. Espitia
Mexico City, Mexico Nutrition and Dietetics School
University of Atlantico
Juan Eduardo René Cortés-Millán Atlantico, Colombia
Laboratorio de Investigación A
Facultad de Nutrición Lucía Famá
Universidad Autónoma del Estado de Morelos Facultad de Ciencias Exactas y Naturales
Cuernavaca, Mexico Departamento de Física, Laboratorio de
Polímeros y Materiales Compuestos (LP&MC)
Maria Jose Costa Instituto de Física de Buenos Aires
Centre of Biological Engineering (IFIBA-CONICET)
University of Minho Universidad de Buenos Aires
Braga, Portugal Buenos Aires, Argentina
CONTRIBUTORS XV

Pablo Fuciños Josef Jampílek

International Iberian Nanotechnology Department of Pharmaceutical Chemistry
Laboratory Faculty of Pharmacy
Braga, Portugal Comenius University
Bratislava, Slovakia
Carlos A. Fuenmayor
Institute of Food Science and Technology (ICTA) Najeh Maissar Khalil
Universidad Nacional de Colombia Pharmaceutical Nanotechnology Laboratory
Bogota, Colombia Department of Pharmacy
Universidade Estadual do Centro-Oeste
Behrouz Ghorani Guarapuava, Brazil
Department of Food Nanotechnology
Research Institute of Food Science Ahmed S. Khan
and Technology College of Engineering and Information
Mashhad, Iran Sciences
DeVry University
Silvia Goyanes Addison, Illinois
Facultad de Ciencias Exactas y Naturales
Departamento de Física, Laboratorio Katarína Kráľová
de Polímeros y Materiales Compuestos Institute of Chemistry
(LP&MC) Faculty of Natural Sciences
Instituto de Física de Buenos Aires Comenius University
(IFIBA-CONICET) Bratislava, Slovakia
Universidad de Buenos Aires
Buenos Aires, Argentina Steva M. Lević
Faculty of Agriculture
Lucas Guz Department of Food Technology and Biochemistry
Facultad de Ciencias Exactas y Naturales University of Belgrade
Departamento de Física, Laboratorio Belgrade, Serbia
de Polímeros y Materiales Compuestos
(LP&MC) Lin Shu Liu
Instituto de Física de Buenos Aires Dairy and Functional Foods Research Unit
(IFIBA-CONICET) Eastern Regional Research Center
Universidad de Buenos Aires Agricultural Research Service
and United States Department of Agriculture
Instituto de Investigación e Ingeniería Wyndmoor, Pennsylvania
Ambiental, Consejo Nacional
de Investigaciones Científicas y Técnicas Alex López-Córdoba
Universidad Nacional de San Martín Facultad de Ciencias Exactas y Naturales
Buenos Aires, Argentina Departamento de Física, Laboratorio
de Polímeros y Materiales Compuestos
(LP&MC)
Humberto Hernández-Sánchez Instituto de Física de Buenos Aires
Escuela Nacional de Ciencias Biológicas del (IFIBA-CONICET)
Instituto Politécnico Nacional Universidad de Buenos Aires
Departamento de Ingeniería Bioquímica Buenos Aires, Argentina
Unidad Profesional Adolfo López Mateos
Mexico City, Mexico
XVI CONTRIBUTORS

Rubén López-Santiago Ollin Celeste Martínez-Ramírez

Departamento de Inmunología Laboratorio de Investigación A
Escuela Nacional de Ciencias Biológicas Facultad de Nutrición
Instituto Politécnico Nacional Universidad Autónoma del Estado de Morelos
Mexico City, Mexico Cuernavaca, Mexico

Mayra Luna-Trujillo Juan Vicente Méndez-Méndez

Laboratorio de Electroquímica y Corrosión Centro de Nanociencias y Micro
Escuela Superior de Ingeniería Química e y Nanotecnologías
Industrias Extractivas Instituto Politécnico Nacional
Instituto Politécnico Nacional Mexico City, Mexico
Mexico City, Mexico
Angélica Gabriela Mendoza-Madrigal
Yun Ma Laboratorio de Investigación A
State Key Laboratory of Plant Genomics Facultad de Nutrición
Institute of Microbiology Universidad Autónoma del Estado de Morelos
Chinese Academy of Sciences Cuernavaca, Mexico
Beijing, China

and Christian Micheletti

European Center for the Sustainable Impact
College of Life Science of Nanotechnology
Tianjin Normal University EcamRicert Srl
Tianjin, China Rovigo, Italy

Miriam Madrid-Mendoza Marilia Moraes-Lovison

Laboratorio de Investigación A Department of Food Engineering
Facultad de Nutrición School of Animal Science and Food Engineering
Universidad Autónoma del Estado de Morelos University of Sao Paulo
Cuernavaca, Mexico Pirassununga, Brazil

Rubiana Mara Mainardes Sara Naji-Tabasi

Pharmaceutical Nanotechnology Laboratory Department of Food Nanotechnology
Department of Pharmacy Research Institute of Food Science
Universidade Estadual do Centro-Oeste and Technology
Guarapuava, Brazil Mashhad, Iran

Arturo Manzo-Robledo Viktor A. Nedović

Laboratorio de Electroquímica y Corrosión Faculty of Agriculture
Escuela Superior de Ingeniería Química e Department of Food Technology and
Industrias Extractivas Biochemistry
Instituto Politécnico Nacional University of Belgrade
Mexico City, Mexico Belgrade, Serbia

Francesco Marra
University of Salerno
Fisciano, Italy
CONTRIBUTORS XVII

Iolanda Olivato Miriam Rodríguez-Esquivel

European Center for the Sustainable Impact Centro de Nanociencias y Micro
of Nanotechnology y Nanotecnologías
EcamRicert Srl Instituto Politécnico Nacional
Rovigo, Italy and
Laboratorio de Oncología Genómica
Semih Ötleş Hospital de Oncología del Centro Médico
Ege University Nacional Siglo XXI
Food Engineering Department Instituto Mexicano del Seguro Social
Izmir, Turkey Mexico City, Mexico

Madelyn Pandorf Marco Roman

School of Sustainable Engineering and the Built European Center for the Sustainable Impact
Environment of Nanotechnology
Arizona State University EcamRicert Srl
Tempe, Arizona Rovigo, Italy

Lorenzo Miguel Pastrana Patricia Rosales-Martínez

International Iberian Nanotechnology Escuela Nacional de Ciencias Biológicas del
Laboratory Instituto Politécnico Nacional
Braga, Portugal Departamento de Ingeniería Bioquímica
Unidad Profesional Adolfo López Mateos
Samantha C. Pinho Mexico City, Mexico
Department of Food Engineering
School of Animal Science and Food Engineering Buket Yalçın Şahyar
University of Sao Paulo Ege University
Pirassununga, Brazil Food Engineering Department
Izmir, Turkey
Radoslava N. Pravilović
Faculty of Technology and Metallurgy Mauricio Salcedo-Vargas
Department of Chemical Engineering Laboratorio de Oncología Genómica
University of Belgrade Hospital de Oncología del Centro Médico
Belgrade, Serbia Nacional Siglo XXI
Instituto Mexicano del Seguro Social
V Ravishankar Rai Mexico City, Mexico
Department of Studies in Microbiology
University of Mysore Fabrizio Sarghini
Mysore, India Biosystem Engineering Section
Department of Agricultural Sciences
Philippe Emmanuel Ramos University of Naples Federico II
Centre of Biological Engineering Naples, Italy
University of Minho
Braga, Portugal
XVIII CONTRIBUTORS

Rajesh P. Shastry Kata T. Trifković

Department of Studies in Microbiology Faculty of Technology and Metallurgy
University of Mysore Department of Chemical Engineering
Mysore, India University of Belgrade
Belgrade, Serbia
Guixue Song
Institute of Marine Science and Technology Alejandra Valdivia-Flores
Shandong University Dirección de Investigación
Shandong, China Secretaria de Investigación y Posgrado
Instituto Politécnico Nacional
Luis Eduardo Suárez-Nájera Mexico City, Mexico
Departamento de Ingeniería Bioquímica
Escuela Nacional de Ciencias Biológicas Jaime Vargas-Cruz
Instituto Politécnico Nacional Departamento de Ingeniería Bioquímica
Mexico City, Mexico Escuela Nacional de Ciencias Biológicas
Instituto Politécnico Nacional
Chin Ping Tan Mexico City, Mexico
Department of Food Technology
Faculty of Food Science and Technology Alonso Villafán-Rangel
Universiti Putra Malaysia Laboratorio de Investigación A
Seri Kembangan, Malaysia Facultad de Nutrición
Universidad Autónoma del Estado de Morelos
Cuernavaca, Mexico
Tai Boon Tan
Department of Food Technology
Paul Westerhoff
Faculty of Food Science and Technology
School of Sustainable Engineering and the Built
Universiti Putra Malaysia
Environment
Seri Kembangan, Malaysia
Arizona State University
Tempe, Arizona
Erik Tedesco
European Center for the Sustainable Impact Oswaldo Ochoa Yepes
of Nanotechnology Facultad de Ciencias Exactas y Naturales
EcamRicert Srl Departamento de Física, Laboratorio
Rovigo, Italy de Polímeros y Materiales Compuestos
(LP&MC)
José António Teixeira Instituto de Física de Buenos Aires
Centre of Biological Engineering (IFIBA-CONICET)
University of Minho Universidad de Buenos Aires
Braga, Portugal Buenos Aires, Argentina

Peggy Tomasula
Dairy and Functional Foods Research Unit
Eastern Regional Research Center
Agricultural Research Service
United States Department of Agriculture
Wyndmoor, Pennsylvania
 PART I

Introduction to Nanotechnology
in the Food Sector
http://taylorandfrancis.com
 CHAPTER 1

Development of Bio-Based Nanostructured Systems

by Electrohydrodynamic Processes

Maria Jose Costa, Philippe Emmanuel Ramos, Pablo Fuciños, José António Teixeira,
Lorenzo Miguel Pastrana, and Miguel Ângelo Cerqueira

CONTENTS

1.1 Introduction .............................................................................................................................. 3

1.2 Electrohydrodynamic Processing ............................................................................................. 5
1.2.1 Equipment and Processing............................................................................................ 5
1.2.2 Parameters..................................................................................................................... 6
1.3 Electrohydrodynamic Processing in the Food Industry ........................................................... 8
1.3.1 Bio-Based Materials Used ............................................................................................ 8
1.3.2 Applications in the Food Industry ................................................................................ 8
1.3.2.1 Encapsulation of Functional Ingredients ...................................................... 10
1.3.2.2 Enzyme Immobilization................................................................................ 11
1.3.2.3 Food Packaging ............................................................................................ 12
1.3.2.4 Bacteriophages.............................................................................................. 13
1.3.2.5 Filtration Membranes.................................................................................... 13
1.4 Future Trends and Final Remarks.......................................................................................... 14
Acknowledgments ........................................................................................................................... 15
References ....................................................................................................................................... 15

1.1 INTRODUCTION

Nanotechnology offers many benefits for the development of structures with enhanced proper-
ties. Due to their dimension, nanostructured systems present different physicochemical properties
(i.e., optical, thermal, absorption, and stability) when compared with micro- and macroscale systems,
making their use in food processing and applications of great interest to the industry. The main aspect
that influences their different behavior at the nanoscale is the area-to-volume ratio. It determines the
physicochemical characteristics and brings several advantages, such as a high encapsulation effi-
ciency and stability, a tailored control release, an enhanced solubility, and the prevention of unde-
sirable chemical reactions (Cerqueira et al. 2014, 2017a).
In recent years, the development of high-performance nanostructured systems for food appli-
cations has improved food safety (e.g., sensor and nanoparticles). These systems have been applied

3
4 NANOTECHNOLOGY APPLICATIONS IN THE FOOD INDUSTRY

in several areas such as the encapsulation and controlled release of compounds, in the development
of new food processing methodologies, and in food packaging. Aiming for commercialization,
several nanotechnology-based products have been market launched in the last 10 years, claiming
enhanced functionality based on their nanosized features (Silva, Cerqueira, and Vicente 2015). The
development of bio-based nanostructured systems, for food applications, has been presented by
researchers through a high number of published works. These works present the potential of
using those systems for several applications such as for the protection and delivery of bioactive
compounds, enzyme immobilization, detection of contaminants and microorganisms through the
development of sensors, removal of chemicals from foods, water purification through nanofiltra-
tion, and the development of nanocomposites for packaging. This great number of applications is
explained by the possibility of developing bio-based nanostructures with distinct architectures that,
according to the materials and methodologies used, can be designed with different shapes and
sizes. The common shapes of bio-based nanostructured systems are spheroids (capsules or particles),
fibers (membranes), thin films (nanocoating for food packaging), and tubes (sensors) (Cerqueira et al.
2017b).
Different approaches can be used for the development of nanostructured systems, giving
importance to the control of the driving forces (e.g., electrostatic and hydrophobic interactions) and
the free energy of the system (e.g., to assess the thermodynamic stability), which leads to systems
with distinct properties and functionalities. During the fabrication process, nanostructured systems
can be developed by the bottom-up and top-down methods, which are two of the common ways to
classify the developed system (Livney 2015; Silva, Cerqueira, and Vicente 2011). Despite all the
advantages of using bio-based nanostructures, the scale-up process from the laboratory to an
industrial scale is still a hard task. The major challenge is controlling the main characteristics and
properties of the nanostructures from the laboratory to the industrial production. In addition, some of
the challenges regarding the scaling-up of nanostructure production are the batch versus continuous
production, the increasing flow rate, and the homogenization process, which leads to a considerable
number of trials for process optimization at industrial scale. While for inorganic nanostructures
(i.e., ZnO, TiO2, and SiO2), the scale-up and commercialization is in the exponential step, where
several companies have shown the capacity to produce several kilograms per day, for organic
nanostructures, the industrial production is still in the beginning (Piccinno et al. 2012; Tsuzuki
2009). Some of the methodologies showing the capacity to be scaled-up for the production of food-
grade bio-based nanostructures are high-pressure homogenization, microfluidics, vibrational atom-
ization (nanospray drier), and nanoprecipitation (Arpagaus 2012; Cerqueira et al. 2014; Lu et al.
2016; Trierweiler and Trierweiler 2011).
Other processes that show promising advantages are the electrohydrodynamic processes, such as
electrospinning and electrospraying that have been explored for the production of food-grade bio-
based nanostructured systems. These new processes are starting to change and improve the food
industry through their use in several applications with a wide range of bio-based materials generally
recognized as safe (GRAS) by the US Food and Drug Administration (FDA) and/or the European
Food Safety Authority (EFSA) (Kaur Bhullar, Kaya, and Jun 2015). Several studies confirmed
that electrospinning and electrospraying processes are very effective in producing bio-based
nanostructures with several advantages that overcome the existing technology limitations, such as
different sizes range (micro to nano) and type of structures (fibers and particles), no heating required,
ability to scale up (multinozzle systems), versatility in morphology (porosity and roughness), and
low or no organic solvent requirement (Ghorani and Tucker 2015). Besides, the use of food-grade
biopolymers for the production of structures able to guarantee high encapsulation efficiencies,
a good stability, and a controlled release of bioactive compounds is also an important advantage
(Anu Bhushani and Anandharamakrishnan 2014). However, they can present some limitations such
as the high price of the equipment and the difficult scale-up of the process (Quirós, Boltes, and Rosal
2016).
DEVELOPMENT OF BIO-BASED NANOSTRUCTURED SYSTEMS 5

1.2 ELECTROHYDRODYNAMIC PROCESSING

In the food industry, electrohydrodynamic processing is still recent and only slightly studied, its
use being unusual in food structuring. Nevertheless, electrospinning and electrospraying are proving
to be excellent options for use in the encapsulation of several functional compounds with interest for
the food industry and in packaging applications (Ghorani and Tucker 2015; Pérez-Masiá, Lagaron,
and López-Rubio 2014). The interest of researchers and industry has been driven by several technical
advantages such as the facility of use, the available equipment, and the possibility of tuning the final
properties of the materials by changing the processing conditions.

1.2.1 Equipment and Processing

The electrohydrodynamic processes use an electrically driven force to produce single or multi-
layer fibers or particles that can be at the micro- and nanometric scale. During the process, an
electrically charged fluid cone jet is formed by the biopolymer solution or molten biopolymer that is
formed between the spinneret and the collector. The applied electric field leads to the formation of a
cone-like shape, called the Taylor cone, that is obtained when the equilibrium of the electric forces
and surface tension is destroyed. In the case of the formation of fibers (electrospinning), continuous
production is achieved when the fluid is ejected from the tip of the Taylor cone and an extended fiber
is formed by an electrostatic field that quickly solidifies on the collector (Figure 1.1a) (Quirós,
Boltes, and Rosal 2016). Due to the high instability of the process, the fiber deposition and orien-
tation is difficult to control, as the fibers are randomly oriented. To achieve a “more” controlled
deposition of the fibers, the movement of the nozzle and/or collector can be controlled as well as the
distance to the collector. For the production of particles (electrospray), the phenomenon is similar
(Figure 1.1b), although, in this case, the electric field causes the deformation and distribution of the
jet into droplets that, after being spray dried, are deposited in the collector (Jaworek and Sobczyk
2008). In the case of electrospraying, it is also possible to electrospray the biopolymers on a cross-
linking solution (mostly used for the alginate cross-linking using CaCl2) to achieve more stable
particles (Ghayempour and Mortazavi 2013).
Choosing a molten biopolymer or the use of a biopolymer solution will highly influence the
equipment requirements for the electrospinning/electrospraying process. While for processing, using
a biopolymer solution, no heating is needed; for the process using a molten biopolymer, equipment
with a heating module is needed. In fact, the use of the melting process is not common in food
applications, mostly because some of the biopolymers and functional compounds used are
thermosensitive and the use of heating can in most cases be a disadvantage (Zhang et al. 2016). One
of the possibilities with electrohydrodynamic processing is to use nozzles with different diameters

Voltage Syringe/pump Voltage Syringe/pump

+ Nozzle/tip + Nozzle/tip

– –

Collector Collector
(a) (b)

Figure 1.1 Main composition of (a) electrospinning and (b) electrospraying equipment during the processing of a
biopolymer solution.
6 NANOTECHNOLOGY APPLICATIONS IN THE FOOD INDUSTRY

and multiaxial geometries (monoaxial, coaxial, or triaxial) that will influence the morphology and the
properties of the produced structures (Chakraborty et al. 2009; Xu et al. 2013). The multiaxial
(coaxial/triaxial) geometries consist in different spinneret components which allow the simultaneous
spinning/spraying of different liquids (Quirós, Boltes, and Rosal 2016). The use of coaxial and
triaxial nozzles allows coating solid, liquid, or gas phases with a solid shell and produce multilayer
structures, bringing several advantages, such as a controlled and tunable release of functional
compounds, and encapsulation of different compounds in the same structure, allowing their release at
different stages.
The collectors can also have different geometries and morphologies, such as flat surface (stainless-
steel plate) or a cylinder (rotating mandrel), that can be in motion during the processing (Cerqueira et
al. 2016; Gómez-Mascaraque, Lagarón, and López-Rubio 2015). The collector movements can be
used to define the pattern of the developed structures (Bu et al. 2012), the use of a flat collector
(working in the x and y directions) and the distance from the spinneret to the collectors (z direction)
allow the development of three-dimensional (3D) structures (Zhang et al. 2016).
Besides the equipment, the different possibilities to electrospin the biopolymer (melted or in
solution) and the processing parameters are also very important factors when the objective is to
obtain fibers or particles. Other important variables are the applied voltage between the nozzle and
collector that, together with the solution and/or biopolymers characteristics, determines the electric
field strength and thus the electrohydrodynamic jetting mode. These parameters will be discussed in
the next section.

1.2.2 Parameters

The electrohydrodynamic processes can be influenced by several parameters that will define
the aspect, size, and morphology of the produced structures. The most influential parameters are
(a) the polymer solution type, (b) processing, and (c) ambient conditions used during the electro-
hydrodynamic process. For the biopolymer solution, specific characteristics such as solvent type,
conductivity, molecular weight, viscosity, and surface tension are the most influential. The process
parameters that influence the obtained structure include the tip to collector distance, diameter of
spinneret, applied voltage, and flow rate. The ambient conditions such as relative humidity, air flow,
and temperature of the surroundings are also important during the process (Chong et al. 2007). The
control of these variables will allow controlling the size, density, and morphology of the produced
structures (Rogina 2014), although it is important to mention that to achieve reproducible results, a
stable cone-jet mode should be obtained (Bock, Dargaville, and Woodruff 2012).
The biopolymer concentration has a direct impact in viscosity, being one of the most important
variables to obtain a stable cone-jet mode. An optimal concentration mainly depends on which
structure type is desired, it being possible to obtain particles or fibers. Normally, with an optimal
concentration range, the low polymer concentrations do not produce a continuous jet, leading to the
production of particles, while for high polymer concentrations, a continuous flow is achieved and
fibers will be formed. In the case of middle term conditions, a mixture of both mentioned structures
will be obtained (i.e., particles and fibers) (López-Rubio and Lagaron 2012; Rogina 2014;
Wongsasulak et al. 2007). During processing, if the flow rate is low or the electric field strength is
high, the Taylor cone cannot be maintained and the process results in a phenomenon such as tilted
jetting, multijetting, and dripping (Zhang et al. 2016).
The electrohydrodynamic process produces dried structures, although to accomplish this, high
concentrations of polymer must be used in order to reduce the solvent content and thus facilitating the
solvent evaporation. However, a low viscosity is needed to allow jet formation; otherwise, a con-
tinuous electrospinning will be not possible (Huang et al. 2003; Shenoy et al. 2005). The solvent used
DEVELOPMENT OF BIO-BASED NANOSTRUCTURED SYSTEMS 7

is an important factor to allow, at first, a proper dissolution of the polymer and, at second, an efficient
drying of the electrospun fibers or electrosprayed particles (Ghorani and Tucker 2015). Therefore,
solvents with lower vapor pressure would permit an easier fiber drying (Wannatong, Sirivat, and
Supaphol 2004). Other works also refer to the fact that the mixture of different solvents might bring
important characteristics such as an easier drying process while maintaining the dissolution of the
polymer (Ghorani, Russell, and Goswami 2013).
One of the factors that influence the polymer solution viscosity the most is the molecular weight
of the polymer. In general, high molecular weight increases viscosity leading to fiber formation (Tao
and Shivkumar 2007). Therefore, low-molecular weight polymers are more likely to produce par-
ticles and high-molecular weight polymers produce fibers with large diameters (Bhardwaj and
Kundu 2010). When the polymer solution is constituted by a mixture of polymers, the molecular
weight of the polymers might also not be so influential in the production of fibers, since the polymers
have enough affinity to create several entanglement points through intermolecular interactions
(Munir et al. 2009).
One of the ways used to understand the capacity of a polymer solution to be electrospun or
electrosprayed is to evaluate the surface tension and conductivity. The surface tension is mainly
dependent on the solvent used rather than on the polymer. High surface tension might prevent the
formation of fibers and capsules or even complicate Taylor’s cone formation (Rogina 2014). A lower
surface tension will allow the utilization of a lower electric field. To avoid a negative influence of
surface tension, a reduction of polymer concentration might be done, although it is important to be
aware that this change will also affect the viscosity of the polymer solution (Moomand and Lim 2015).
The conductivity of the solution is influenced by the polymer, solvent type, and the presence of salts.
More conductive solutions produce smaller-diameter fibers through electrospinning (Zong et al. 2002;
Jiang et al. 2004). Therefore, with the same conditions, a more conductive solution will produce a
higher elongated jet, thus producing a fiber with a smaller diameter (Tan et al. 2005). Aiming at
controlling this parameters, it is common to use inorganic salts, which are added to increase electrical
conductivity due to their smaller ionic diameter and higher ion mobility (Su et al. 2011).
Regarding the parameters that are controllable in the equipment used, the applied voltage, flow
rate, and collector distance are the most studied. The flow rate controls the amount of polymer that
flows to the collector during a certain period of time. The flow rate should respect the drying time, to
create dried fibers or particles. An increase in the flow rate will generate structures with bigger
diameters, as a consequence of a larger amount of electrospun polymer (Zong et al. 2002).
The applied voltage controls the electric field strength created between the spinneret and the
collection point. Ghorani and Tucker (2015) worked to overcome the normal behavior of a dropwise
solution to accomplish a Taylor’s cone. It is extremely dependent on parameters such as flow rate,
surface tension, and viscosity. Higher voltages will increase the velocity of the jet and, as a con-
sequence, the transported solution volume (Zong et al. 2002). Usually, there is a voltage range able to
produce the minimal average fiber diameter, where a higher or lower voltage will increase the
average fiber diameter (Ghorani and Tucker 2015). The distance between the tip and the collector is
essential to allow fibers and particles to dry and should be adjusted according to the polymer con-
centration and applied voltage (Guo et al. 2013; Zou et al. 2011). In general, a greater distance
between the tip and the collector will create longer flight times and higher solvent evaporation.
Consequently, the diameter of the produced fibers will be smaller and the tendency for particles
creation will be reduced (Lin et al. 2008; Yuan et al. 2004).
Other parameters that should be controlled are the ambient conditions during processing.
Temperature is an ambient parameter that mainly affects the polymer viscosity and the fiber drying
process. Higher temperatures would create a lower polymer viscosity, and as a consequence, the fiber
diameters will be smaller (Bhardwaj and Kundu 2010). Humidity can influence the fiber drying
process, where a low humidity allows an easier fiber drying process (Demir et al. 2002).
8 NANOTECHNOLOGY APPLICATIONS IN THE FOOD INDUSTRY

1.3 ELECTROHYDRODYNAMIC PROCESSING IN THE FOOD INDUSTRY

The use of bio-based materials in the development and production of nanostructures through
electrohydrodynamic processing is revealing several structural and functional advantages. Regarding
commercial applications in the food industry, the use of biopolymers to produce nanofibers and
nanoparticles, using electrohydrodynamic processes, has a wide area of applications such as new
food ingredients, food additives encapsulation, high-performance (active) food packaging and
coatings, and biosensors. However, and at this moment, only few companies are able to produce it
using biopolymers at industrial scale (one of the cases is Bioinicia 2017 in Spain). Another kind of
application with high potential is the use of electrohydrodynamic processing in the confectionery
industry since the electrospun structures allow the creation of new textures and coatings in foods,
leading to new and enhanced organoleptic properties (e.g., chocolate) (Altay and Okutan 2015; Luo
et al. 2012a; Nieuwland et al. 2014).
In this section, the bio-based materials used in electrohydrodynamic processing will be explored,
as well the food applications where they can be used.

1.3.1 Bio-Based Materials Used

Several bio-based materials from diverse origins can be used in electrohydrodynamic processes.
Bio-based materials can be divided into the following:
• Bio-based materials extracted from biomass, such as polysaccharides and proteins
• Bio-based materials obtained from biomass-derived monomers where the final biodegradable polymer is
obtained through classical chemical synthetic routes, such as poly(lactic)acid (PLA)
• Polymers obtained from natural or genetically modified microorganisms, like xanthan gum and
polyhydroxyalkanoates

Bio-based materials are all suitable for electrospinning and electrospraying with different purposes
such as encapsulation of bioactive compounds, food packaging and coating, enzyme immobilization,
and production of filtration membranes (Anu Bhushani and Anandharamakrishnan 2014; Fabra,
López-Rubio, and Lagaron 2016; Ghorani and Tucker 2015). Bio-based materials are also promising
choices due to their biodegradability and biocompatibility characteristics and to the fact that they can
be tailored to the desired properties turning them into versatile carriers (Aceituno-Medina et al. 2014;
Nieuwland et al. 2014; Stijnman, Bodnar, and Hans Tromp 2011). Another advantage of these
materials is the fact that they can be used simultaneously with different methods (e.g., layer-by-layer,
cross-linking) to enhance the final properties of structures. Concerning regulatory aspects, the choice
will fall in materials generally recognized as safe (GRAS) and approved by the regulatory agencies
(i.e., EFSA and FDA). From an economic point of view, elements such as a continuous supply, cost,
and simplicity of use are all taken into account for the selection of the matrix source material. Some of
the biopolymers used for electrospinning and/or electrospraying are presented in Table 1.1.

1.3.2 Applications in the Food Industry

The main objectives of the food industry is to ensure the stability and quality of the food products
as well the process efficiency. It seeks to achieve these demands and, at the same time, maintain food
processing as simple as possible. So new and innovative food processing technologies play an
important role in the continuous improvement of food processing and their products.
This is the case for new processing technologies such as the electrohydrodynamic process that can
help the food industry in achieving greater performance, higher versatility, and innovative approaches
to the different types of applications. Some of these applications are the encapsulation of bioactive
compounds, food packaging and coating, enzyme immobilization, and bacteriophage incorporation
(Table 1.2) (Fabra, López-Rubio, and Lagaron 2016; Lee et al. 2016; Wen et al. 2016).
DEVELOPMENT OF BIO-BASED NANOSTRUCTURED SYSTEMS 9

Table 1.1 Bio-Based Material Source and Type Used in Electrohydrodynamic Processing, Developed
Structure, and Encapsulated Compounds
Biomaterial Biomaterial Encapsulated
Source Type Structure Compounds Reference
Directly extracted Chitosan Electrospun fibers CNMA Rieger and Schiffman
from biomass (2014)
Chitosan Particles and fibers (−)-Epigallocatechin Gómez-Mascaraque,
gallate Sanchez, and López-
Rubio (2016)
Sodium alginateElectrospray Folic acid Bakhshi et al. (2013)
particles
Starch Electrospun fibers BCNW Fabra et al. (2016)
Alyssum Electrospray D-limonene Khoshakhlagh et al.
homolocarpum particles (2017)
seed gum
WPI Electrospun fibers α-tocopherol Fabra, López-Rubio,
and Lagaron (2016)
Whey protein Electrospray Folic acid Pérez-Masiá et al.
concentrate particles (2015)
Caseinate Electrospun fibers N/A Tomasula et al. (2016)
SPI Electrospun fibers Allyl isothiocyanate Vega-Lugo and Lim
(2009)
API Electrospun fibers Nisin Soto et al. (2016)
Zein Coaxial Rose hip seed oil Yao et al. (2016)
electrospinning
fibers
Electrospray Green tea catechins Anu Bhushani, Kurrey,
particles and
Anandharamakrishnan
(2017)
Electrospun CNMA Cerqueira et al. (2016)
interlayers fibers
Zein Electrospun fibres β-carotene Fernandez, Torres-
Giner, and Lagaron
(2009)
Gelatin Electrospray Polyphenols Gómez-Mascaraque,
particles Lagarón, and López-
Rubio (2015)
Gelatin Electrospun fibrils N/A Nieuwland et al. (2014)
Oil-based/biomass- Polycaprolactone Electrospun fibers N/A Martins et al. (2015)
derived monomers PVA Electrospun fibers Eugenol Kayaci, Ertas, and Uyar
(2013)
PEO Electrospun fibers N/A Sullivan et al. (2014)
PLA Coaxial N/A Nguyen, Chung, and
electrospinning Park (2011)
fibers
Polymers produced PHBV Electrospun fibers Zinc oxide Castro-Mayorga et al.
by natural or (2017)
genetically Kefiran Electrospun fibers N/A Esnaashari et al. (2014)
modified
Pullulan Electro-wet-spinning N/A Kong and Ziegler (2014)
microorganisms
fibers
10 NANOTECHNOLOGY APPLICATIONS IN THE FOOD INDUSTRY

Table 1.2 Examples of Applications of Electrohydrodynamic Processes in the Food Industry

Application Feature Technique Notes Reference
Encapsulation Functional Electrospinning Encapsulation of López-Rubio et al.
ingredient Bifidobacterium (2012)
animalis subsp. Lactis
Bb12 strains using WPC
and pullulan
Encapsulation Functional Electrospraying Encapsulation of folic acid Bakhshi et al. (2013)
ingredient
Enzyme Biosensor Electrospinning Glucose oxidase in PVA Ren et al. (2006)
immobilization
Food packaging and Active multilayer Multilayer Multilayer structure with Cerqueira et al.
coating bio-based electrospinning PHBV 8% and zein as (2016)
antimicrobial interlayer with CNMA
packaging
Bacteriophages Antimicrobial Coaxial T4 bacteriophage fibers Korehei and Kadla
incorporation packaging electrospinning using PEO (2013)
New textures Protein-based Electrospinning Gelatin and zein Nieuwland et al.
production textures electrospinning (2014)

1.3.2.1 Encapsulation of Functional Ingredients

The use of nanostructures for the encapsulation of functional ingredients has been presented as
one of the most interesting systems for the protection, delivery, and controlled release of bioactive
compounds and probiotics. It is known that functional ingredients can be very sensitive to different
conditions (light, temperature, and pH) (Altay and Okutan 2015). Moreover, the lipophilic or
hydrophilic nature of bioactive compounds can influence their behavior during the encapsulation
process. So the use of bio-based materials for functional ingredient incorporation is a subject of
special attention due to many applications with different objectives, namely, the following:

• Controlled release (Jiang, Wang, and Zhu 2014)

• Control temperature influence (Camerlo et al. 2014)
• Functional activity maintenance (Cerqueira et al. 2016)

Encapsulation arises as a controlled delivery system where stability and functional performance
will depend on their internal and external morphologies. The encapsulation of the functional ingre-
dients in foods must ensure no undesirable impact on the organoleptic properties. At the same time, it
must maintain their physicochemical properties providing a controlled release, ensuring the activity of
the bioactive component when eaten. The major benefits of encapsulation come from small dimension
matrices with high surface/volume ratios. These factors will lead to higher solubility, better aggre-
gation stability, faster diffusion rates, and an increase in bioavailability (Davidov-Pardo, Joye, and
McClements 2015).
Several methods were already used and evaluated to produce encapsulation structures. Conven-
tional methods such as coacervation (Yang et al. 2015), spray drying (Pérez-Masiá et al. 2015),
nanoemulsion (Silva, Cerqueira, and Vicente 2012), and extrusion (Ramos et al. 2017) are commonly
used for encapsulation. Spray drying is one of the most commonly used in the food industry due to its
affordable operational costs. However, traditional methods can present some disadvantages such as the
loss of volatile compounds and of probiotic bacteria viability due to the temperature needed during the
process. Electrohydrodynamic processes appear as a novel method to encapsulate bioactive compo-
nents due to their easy-to-use characteristics and advantages such as one-step formulation and the lack
of need to use organic solvents or high temperature. Many studies were already performed and proved
DEVELOPMENT OF BIO-BASED NANOSTRUCTURED SYSTEMS 11

5 µm
50.0 µm
(a) (b)

Figure 1.2 Scanning electron microscopy images of (a) WPC capsules obtained through electrospraying from
40 wt.% WPC aqueous solutions and (b) zein fibers obtained through electrospinning from 33 wt.%
zein ethanolic solutions. (Reprinted from Innovative Food Science and Emerging Technologies,
13, A. López-Rubio and J. M. Lagaron, Whey protein capsules obtained through electrospraying
for the encapsulation of bioactives, 200–6, Copyright (2012), with permission from Elsevier; with
kind permission from Springer Science+Business Media: Food and Bioprocess Technology, Use
of electrospinning to develop antimicrobial biodegradable multilayer systems: Encapsulation of
cinnamaldehyde and their physicochemical characterization, 9, 2016, 1874–84, M. A. Cerqueira et al.)

that encapsulation by electrospinning/electrospraying is very effective when the aim is the encapsu-
lation of bioactive compounds. Aceituno-Medina et al. (2014) tested the encapsulation of folic acid in
amaranth protein isolate (API) and pullulan electrospun fibers and observed an increase in the thermal
stability of the structure, assessed by the photoprotection during 120 min, avoiding folic acid degra-
dation. Using whey protein concentrate (WPC), López-Rubio and Lagaron (2012) showed the pos-
sibility of using the electrospray process to encapsulate b-carotene in nanoparticles with a high
encapsulation efficiency and, at the same time, stabilize b-carotene against photooxidation. These
results showed a possible solution for the use of photosensitive bioactive compounds in food appli-
cations. López-Rubio and Lagaron (2012) used WPC to encapsulate Bifidobacterium animalis subsp.
lactis Bb12 in electrospray particles. The results showed a substantial increase in the viability of the
Bifidobacterium strain at 20°C and a greater protection, even at high relative humidity, showing
conditions for successful functional food applications. Figure 1.2 shows some of the structures
developed for the encapsulation of food compounds. Figure 1.2a and b shows nanocapsules and
nanofibers produced through electrohydrodynamic processing using WPC and zein, respectively.

1.3.2.2 Enzyme Immobilization

Enzymes are natural catalysts that have high regio- and stereoselectivity and enhance the reaction
rate of chemical reactions in living organisms. They are expensive, very sensitive, and unstable,
leading to limitations when they are used in industry (Anu Bhushani and Anandharamakrishnan
2014; Lee et al. 2016). The immobilization process can solve some of these limitations, confining the
enzyme in a specific region to overcome the enzyme sensitivity and instability problems and to allow
a possible future reuse (Lee et al. 2016). Several techniques such as entrapment, cross-linking,
adsorption, and covalent bonding are used to immobilize enzymes. The most used technique is
covalent bonding, which allows the reuse of the enzyme and, because of it, is one of the methods that
present the most robust immobilization. However, the type of support (chemical and morphological
structure) in covalent bonding significantly influences the immobilized enzyme activity. To over-
come these challenges and ensure a high surface area (to increase the functional groups for covalent
bonding) and low diffusion limitations (to substrate and product), nanofibers produced by
12 NANOTECHNOLOGY APPLICATIONS IN THE FOOD INDUSTRY

electrospinning appear as a solution due to their high surface area, open pore structure, and large
lateral interspace between nanofibers (Lee et al. 2016). Liu et al. (2012) tested the glucose oxidase
immobilization in poly(vinyl) alcohol (PVA)/chitosan/tea extract and observed the antibacterial
activity against Escherichia coli and Staphylococcus aureus. This system exhibited 73% of deoxi-
dization in tested samples and exerted inhibition of the growth of microorganisms, and so is suitable
as a novel food packaging material able to extend the food shelf life. Several enzymes have a crucial
importance in food industries; some studies have already shown an improvement in the efficiency
and catalyzing capability of immobilized enzymes. For instance, Lee et al. (2016) immobilized lipase
via cross-linking in methylcellulose nanofibrous mat where more than 90% of the initial activity was
maintained after seven reuses. These results are very important for food applications since lipases are
used in bioconversion reactions in several food industries, such as the bakery and dairy industries.

1.3.2.3 Food Packaging

Food packaging, with films and coatings, are very important since they protect food along the
food chain and guarantee the quality and safety of food products. Besides protection, they can also
be used to provide extra functionalities such as antioxidant and antimicrobial activities or used as
carriers of probiotics and nutrients. The use of electrospinning for food active packaging pro-
duction is a very promising technique since it allows the production of nanolayered structures with
controlled release ability by different triggers (relative humidity, temperature, and pH). Another
interesting application is the use of electrospinning to produce food packaging with biosensors.
This technology will create a response depending on the internal and external environment pro-
viding information about food quality and safety (Ren et al. 2006). One example is described by
Luo et al. (2012b) that produced an immunobiosensor using magnetic nanoparticles immobilized in
cellulose nitrate nanofibers, produced by electrospinning. This structure was conjugated with an
antibody specific for E. coli O157:H7, allowing observation that the biosensor was capable of
detecting E. coli at a low level (67 colony-forming units/mL in a total detection time of 8 min).
These results showed that this is a sensitive, reliable, fast, and low-cost biosensor, which can be
used in food applications.
Regarding food coatings, this application is very important due to the barrier properties that can
lead to the creation of better packaging materials. A possible methodology to apply a coating in
food is spray coating. In this case, the electrospray is used to form a coating in the food product, it
being possible to apply a uniform coating with a controlled deposition rate and film thickness.
A major advantage is the ability to control the droplet size through the adjustment of the voltage
and flow rate of the instrument and the possibility to control the spraying solution concentration
(Anu Bhushani and Anandharamakrishnan 2014). Yao et al. (2016) encapsulated rose hip seed oil
in zein prolamine fibers through coaxial electrospinning and showed that these compounds have a
promising food packaging potential since they delayed the degradation of fruits (bananas and
kumquats) for 7 days.
The electrohydrodynamic process can also be used as a methodology for the development of
interlayers for multilayer food packaging. The electrospun material can be used as a natural adhesive
that ties together different biopolymer layers and, at the same time, improves biopolymer barrier
properties. These advantages are highly important in food packaging/coating since the adhesion of
different materials (i.e., hydrophobic with hydrophilic) is very important in multilayer food packaging.
Barrier properties are also very important to ensure food quality and will depend on the type of food
product (e.g., fresh or dry food product) and their different demands (e.g., high or low barrier).
Interlayer methodology presents several advantages, making this methodology very useful for food
packaging since it is a natural adhesive method, improves the physical performance (optimizing
coating barrier properties), and allows the interlayers to be tailor made based on the final objective.
Some biopolymers such as soy protein (Vega-Lugo and Lim 2009) and zein (Cerqueira et al. 2016)
DEVELOPMENT OF BIO-BASED NANOSTRUCTURED SYSTEMS 13

were already used and proved the possibility of using this methodology to assemble aliphatic polyester
with natural biopolymers (Fabra et al. 2013). Recently, Cerqueira et al. (2016) tested a multilayer
system using poly(3-hydroxybutyrate-co-3-hydroxyvalerate 8 (PHBV8) (8% valerate content) and
zein with and without cinnamaldehyde (CNMA) as interlayer. To complete this system, different outer
layers were tested (i.e., PHBV8 and alginate-based films). The results showed antibacterial activity for
Listeria monocytogenes to be more effective with PHBV8 with CNMA-loaded zein and using PHBV8
as outer layer. These results revealed a methodology and a system with promising characteristics for
active bio-based food packaging.
Other studies were done with different matrices. For instance, Fabra, López-Rubio, and Lagaron
(2016) used electrospun fibers as a coating layer (whey protein isolate [WPI], zein, and soy protein
isolate [SPI]) in wheat gluten films, forming a bilayer biostructure to encapsulate alpha-tocopherol.
The release profiles and stability of alpha-tocopherol in a sterilization process were evaluated. Zein
matrix revealed the best results with higher stability and slower release due to a more efficient water
vapor reduction. These results lead to a new method for the creation of bioactive packaging systems
for food application. In addition, some tests with nanocomposites and multilayered design were done
and evaluated by Fabra et al. (2016) that used thermoplastic corn starch bionanocomposites and 15%
bacterial cellulose nanowhiskers (BCNWs), coated with electrospun poly(3-hydroxybutyrate)–
BCNW fibers. This system showed an improvement in water vapor permeability with a reduction of
70% and an enhanced barrier performance since it reduced the oxygen permeability by 98%, which
are very important features in food applications.
All these functionalities are very important for the food industry leading to the creation of
innovative active food packaging, with optimized and desired properties (protection, controlled
release, sensors, flavoring, antimicrobial, and probiotic) and to boost sustainability.

1.3.2.4 Bacteriophages

Bacteriophages (or phages) are viruses that infect and kill only a specific bacterium, being used to
ensure food safety and quality. Phages can be used for specific bacteria control in different steps of
food production and storage, although with antibiotic technology, appearance phage therapy was left
aside. The application of phages to reduce the concentration of bacterial pathogens is very promising to
ensure food preservation (Korehei and Kadla 2013). Techniques such as electrospraying and
electrospinning are suitable to incorporate phage and maintain their activity since electrospun materials
are very flexible. Nowadays, there are some works focused on the incorporation of phages using these
techniques, although more studies are needed in the near future. Using electrospinning/electrospraying
in phage suspensions leads to phage deactivation due to the fast water evaporation during single-nozzle
electrospinning. Coaxial electrospinning was successful to encapsulate T4 phage by placing it in the
core of the fiber. Regarding the release profile, using an optimized system with higher-molecular
weight polyethylene oxide (PEO) or blending with hydrophobic cellulose in aqueous buffer medium
allowed a slower release rate (Korehei and Kadla 2013).

1.3.2.5 Filtration Membranes

One of the interesting applications in food industries is the use of electrospun fibers for liquid
filtration where it is needed to remove some contaminants, such as metals, and where the conven-
tional purification methods are inefficient. One of the ways is the development of functionalized
biopolymer-based nanofiber membranes able to retain toxic heavy metal ions, such as arsenate, and
therefore obtain a clean water that can be used in food processes (Min et al. 2016). Another inter-
esting application is the use of nanofiber filter membranes as antimicrobial filter. One of the studies
was performed by Desai et al. (2009), which presented a chitosan-based nanofiber membrane with a
capacity to bind to hexavalent chromium and a reduction of up to 2–3 log of E. coli bacteria.
14 NANOTECHNOLOGY APPLICATIONS IN THE FOOD INDUSTRY

1.4 FUTURE TRENDS AND FINAL REMARKS

The increasing number of publications on the use of electrohydrodynamic processing reveals the
interest in the potential applications of this technology in the food industry. This trend is confirmed
by the number of companies that are selling equipment and devices for the production of fibers and
particles using electrohydrodynamic processing. The scale-up of the technology using bio-based
materials that present GRAS status is still under development when compared with petroleum-/
synthetic-based ones. This is mainly due to the low variability of the materials and high availability
when compared with bio-based biopolymers extracted from renewable sources. The injection system
has to be modified when the process is scaled-up. Thus, multispinneret systems arranged either in
uniaxial configuration or in circular geometry are used in equipment at industrial scale. One of the
possibilities is the use of a hemispherical cap above the nozzle, which was shown to be useful to
increase the flow rate (from 2 to 65 mL/h) during the electrospraying of ethanol, maintaining the
stability of the cone-jet (Morad et al. 2016). The possibility to produce nanofibers and nanoparticles
at the industrial scale using different pieces of equipment such as Fluidnatek™ (Bionica 2017) and
Nanospider™ (Elmarco 2016) presented by companies and researchers (Kim et al. 2015); and the
number of patent applications shows the interest in the use of this technology in the food industry. In
fact, since the first patent in 1934 (Formhals 1934) regarding the production of fibers using cellulose
derivatives by electrohydrodynamic processing, a great number of patents have been granted in this
topic for several applications, although only few of them have been applied regarding the use of this
technology for food purposes. Recent patent applications claim the possibility of producing particles
and fibers using protein and polysaccharides mixtures. In 2012, Pepsico, Inc. presented an inven-
tion where conjugated proteins are produced using electrospinning (e.g., they prepared dextran-
conjugated whey proteins) where the main advantages over other methods for protein conjugation
are that the electrospinning allows the glycosylation in shorter annealing times and with a higher
yield. Also, an invention presenting the production methods of micro- and nanostructures using
amaranth protein combined with a biopolymer was presented. The invention describes the use of
electrospinning, electrospray, or bow-spinning for the production of the structures (López-Rúbio
et al. 2015). The same authors presented an invention for the protection of biological material in
general and thermolabile substances (i.e., microorganisms and viruses) that requires protection to
extend their shelf life or broaden the scope of applications. This patent shows how to encapsulate the
microorganisms Lactobacillus plantarum and Bifidobacterium longum subsp. infantis (CECT 4552)
with increasing viability by electrohydrodynamic processing, protecting the microorganisms against
stress conditions such as temperature, relative humidity, or pH (Lagarón, Pérez-Másia, and López-
Rubio 2015).
Therefore, in the next years, it is expected to find structures produced using electrohydrodynamic
processes in several food products and systems (packaging), as well as during their processing
(filtration). It is also expected that their use in packaging materials and food processing (filtration and
enzyme immobilization) will be achieved in consumer goods fairly shortly, while encapsulation
using nanoparticles and nanofibers for delivery and controlled release applications will need further
safety studies. These studies should show how these systems will protect and release compounds
with different characteristics and how they will behave in the gastrointestinal tract, in order to
guarantee safety and acceptance among regulatory agencies, industry, and consumers. One of the
applications that can bring huge benefits to the food industry in the development of new and
innovative foods is the use of electrohydrodynamic processing for 3D printing foods (Zhang et al.
2016). New foods (soft, palatable, and nutritious) can be created through the deposition of polymer
fibers and particles. They can help to support the development of foods for consumers needing
special foods (e.g., elderly people) due to physiological dysfunctions (e.g., dysphagia) and/or spe-
cific nutritional requirements (e.g., protein, iron, and calcium) (Aguilera and Park 2016). Some
studies showed the possibility of using electrohydrodynamic processing to produce electrospun food
DEVELOPMENT OF BIO-BASED NANOSTRUCTURED SYSTEMS 15

structures, such as chocolate and cocoa coatings (using the heated process) (Gorty and Barringer
2011; Marthina and Barringer 2012). Also some studies were performed to produce a meat replacer
by using gelatin and zein nanofibers (Nieuwland et al. 2014).
One of the key points of using innovative technologies and submicro- and nanomaterials in
foods, besides the legislation, is the consumer acceptance. In fact, it is the consumers who will decide
wether electrohydrodynamic processed products will reach the market and if they will be widely
consumed.

ACKNOWLEDGMENTS

This study was supported by the Portuguese Foundation for Science and Technology (FCT)
under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020
(POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145-FEDER-000004)
funded by the European Regional Development Fund under the scope of Norte2020—Programa
Operacional Regional do Norte.
M. J. Costa is the recipient of a fellowship supported by a doctoral advanced training (call
NORTE-69-2015-15) funded by the European Social Fund under the scope of Norte2020—
Programa Operacional Regional do Norte.
This research was supported by Norte Regional Operational Program 2014–2020 (Norte2020)
through the European Regional Development Fund (ERDF) Nanotechnology-based functional
solutions (NORTE-01-0145-FEDER-000019).

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i n a t e + L . c a s e i + m u c i l a g e ns R o u g h n
e s s ( n m ) (d) (e)

(a) (b) (c)

5 µm 4 3 2 1 5 µm 4 3 2 1 5 µm 4 3 2 1

5 µm 4 3 2 1

5 µm 1 2 3 4 5 µm 1 1 2 3 4 5 µm 1 2 3 4 5 µm 2 3 4 5 µm 1
2 3 4 5 µm 5 µm 4 3 2 1 5 µm 4 3 2 1

Figure 17.9 AFM images of (a) alginate microspheres, (b)

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