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NANOMATERIALS
FOR FOOD
APPLICATIONS
This page intentionally left blank
NANOMATERIALS
FOR FOOD
APPLICATIONS
Edited by

AMPARO LOPEZ RUBIO
Preservation and Food Safety Technologies Department, Institute of
Agrochemistry and Food Technology (IATA-CSIC), Valencia, Spain
FABRA ROVIRA
MARIA JOSE
MARTA MARTINEZ SANZ


LAURA GOMEZ
GOMEZ-MASCARAQUE
Food Chemistry and Technology Department, Teagasc Food
Research Centre, Moorepark, Ireland
Elsevier
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To my beloved uncle Vicente Rubio and to all the researchers, like him,
who show a true passion for advancing scientific knowledge.
CONTENTS
Contributors xiii
Preface xvii
1. Nanomaterials for Food Applications: General Introduction and Overview

of the Book 1
pez Rubio, Laura G. Go
Amparo Lo mez-Mascaraque, M.J. Fabra, M. Martínez Sanz
1.1 Introduction 1
1.2 Potential of Nanotechnologies to Improve Consumer Health 3
1.3 Nanotechnologies for Food Processing and Sensing Applications 4
1.4 Food Packaging Nanotechnologies 6
1.5 Characterization, Toxicological Assessment, and Regulatory Status of Nanomaterials
in Food 7
References 8
Part 1: Nanoingredients
2. Nanostructured Systems to Increase Bioavailability of Food Ingredients 13
María Artiga-Artigas, Isabel Odriozola-Serrano, Gemma Oms-Oliu, Olga Martín-Belloso
2.1 Introduction 13
2.2 Nanostructured Delivery Systems to Encapsulate Bioactive Compounds 14
2.3 Bioaccessibility and Bioavailability From Nanoencapsulation Systems 19
2.4 Industrial Applications 24
2.5 Future Trends and Concluding Remarks 27
References 28
Further Reading 33
3. Nanoencapsulation: Techniques and Developments for Food Applications 35

Elham Assadpour, Seid Mahdi Jafari
3.1 Introduction 35
3.2 Nanoencapsulation Versus Microencapsulation 36
3.3 Classification of Nanoencapsulation Techniques 37
3.4 Gastrointestinal Fate and Bioavailability of Nanoencapsulated Food Components 53
3.5 Conclusion and Final Remarks 53
References 55
vii
viii Contents
4. Nanostructured Minerals and Vitamins for Food Fortification and Food

Supplementation 63
Jesper T.N. Knijnenburg, Lidija Posavec, Alexandra Teleki
4.1 Introduction 64
4.2 Synthesis Methods of Nanoparticles for Food Applications 65
4.3 In Vitro and In Vivo Assessment 68
4.4 Latest Developments in Research 76
4.5 Critical Evaluation of Nanostructured Food Fortificants and Supplements 83
4.6 Conclusions and Future Trends 84
References 85
Part 2: Nanotechnologies for Food Processing

5. Recent Developments in Nanofiltration for Food Applications 101
A.W. Mohammad, Y.H. Teow, K.C. Ho, N.A. Rosnan
5.1 Introduction 101
5.2 Theory and Mechanism of Nanofiltration Membranes 102
5.3 Applications in Food Industry 105
5.4 Issues and Challenges 115
List of Abbreviations 116
References 117
Part 3: Nanosensors for Food Quality and Safety

6. Nanoparticle-Based Aptasensors for Food Contaminant Detection 123
Richa Sharma, K.S.M.S. Raghavarao
6.2 Transduction Approaches for Nanoparticle-Based Aptasensing 128
6.3 Conclusions and Future Perspective 138
Abbreviations 139
Acknowledgments 140
References 140
Further Reading 144
Contents ix
7. Nanoparticles as Biosensors for Food Quality and Safety Assessment 147

K.V. Ragavan, Suresh Neethirajan
7.2 Components of Biosensor and Their Properties 149
7.3 Nanoparticles-Based BiosensorsdClassification 154
7.4 Semiconductor Quantum Dots 184
7.5 Perspectives and Conclusions 193
Acknowledgments 194
References 195
Part 4: Nanotechnologies for Food Packaging

and Biopackaging
8. Nanotechnology in Food Packaging 205
Pilar Hernandez-Mun
~oz, Josep Pasqual Cerisuelo, Irene Domínguez,
pez-Carballo, Ramo
Gracia Lo n Catala, Rafael Gavara

8.2 Nanomaterials Used in the Development of Polymer Nanocomposites 206
8.3 Applications of Nanocomposites in Food Packaging 211
8.4 Human Health Risks and Environmental Impact of Nanotechnology Applied
to Food Packaging 223
8.5 Regulations of Nanotechnology in the Food Sector 226
8.6 Concluding Remarks 227
References 227
Further Reading 232
9. Bioactive Packaging: Combining Nanotechnologies With Packaging

for Improved Food Functionality 233
Pablo R. Salgado, Luciana Di Giorgio, Yanina S. Musso, Adriana N. Mauri
9.1 Requirements for Food Packaging: From Traditional Packaging to Bioactive Ones 233
9.2 Bioactive Compounds for the Formulation of Bioactive Packaging 236
9.3 Development of Nano-Sized Carrier Systems for Bioactive Compounds 243
9.4 Effect of Nanostructures on Their Passage Through the Gastrointestinal
Tract and Bioavailability 255
x Contents
9.5 Bioactive Packaging Containing Nanostructures 256

9.6 Conclusions and Future Trends 259
Acknowledgments 260
References 260
Part 5: Nanotoxicology and Regulatory Status

10. Analytical Challenges and Practical Solutions for Enforcing Labeling
of Nanoingredients in Food Products in the European Union 273
Manuel Correia, Eveline Verleysen, Katrin Loeschner

10.2 Potential Sources of Nanoparticles in Food 275
10.3 The Issue of Sample Preparation 282
10.4 Analytical Methods for Inorganic Nanoparticles in FooddPros and Cons 287
10.5 Suggestion for a Screening Strategy for Inorganic Nanoparticles in Food 295
10.6 The Next ChallengedAnalysis of Organic and Pure Carbon-Based Particulate
Nanomaterials in Food 296
10.7 Validation Studies and Reference Materials for Nanoparticles in Food 299
10.8 Conclusions 302
Acknowledgments 303
References 303
11. Characterization of Nanomaterials: Tools and Challenges 313

Thilak Mudalige, Haiou Qu, Desiree Van Haute, Siyam M. Ansar, Angel Paredes,
Taylor Ingle
11.2 Challenges of Nanoparticle Analysis in Food Matrices 315
11.3 Size and Shape Analysis 318
11.4 Size Separation Techniques 328
11.5 Crystal Structure 337
11.6 Elemental Composition Analysis 337
11.7 Characterization of Surface Chemistry of Nanomaterials 340
11.8 Summary 343
Abbreviations 344
Acknowledgments 344
References 345
Contents xi
12. Toxicological Hazard Analysis of Nanomaterials With Potential for

Utilization in Consumer Goods 355
Ali Kermanizadeh, David M. Brown, Peter Møller
12.2 Physicochemical Characteristics of Nanomaterials Which Govern Potential Toxicity 356
12.3 Exposure Routes of Nanomaterials 359
12.4 Summary and Conclusions 369
12.5 Current and Future Challenges 371
Conflict of Interest Statement 373
References 373
Further Reading 380
13. Regulatory Status of Nanotechnologies in Food in the EU 381

Kirsten Rasmussen, Hubert Rauscher, Stefania Gottardo, Eddo Hoekstra,
Reinhilde Schoonjans, Ruud Peters, Karin Aschberger
13.2 Regulatory Definitions of Nanomaterial in the European Union 395
13.3 Overview of EU Legislation Relevant to Food and Nanomaterials 396
13.4 EU Agencies and Laboratories and the Evaluation Process 401
13.5 EFSA’s Preparation for Safety Assesment of Nanomaterials 404
13.6 Conclusions 405
13.7 Disclaimer 406
Acknowledgments 406
References 407
Index 411
CONTRIBUTORS
Siyam M. Ansar
Office of Regulatory Affairs, Arkansas Laboratory, U.S. Food and Drug Administration, Jefferson,
AR, United States
María Artiga-Artigas
Department of Food Technology, University of Lleida e Agrotecnio Center, Lleida, Spain
Karin Aschberger
European Commission’s Joint Research Centre, Ispra, Italy
Elham Assadpour
Department of Food Science and Technology, Baharan Institute of Higher Education, Gorgan,
Iran
David M. Brown
Heriot Watt University, School of Engineering and Physical Sciences, Nano Safety Research
Group, Edinburgh, United Kingdom
Ram on Catala
Institute of Agrochemistry and Food Technology, IATA-CSIC, Paterna, Spain
Josep Pasqual Cerisuelo
Manuel Correia
Technical University of Denmark, National Food Institute, Division of Food Technology,
Lyngby, Denmark
Luciana Di Giorgio
Centro de Investigaci
on y Desarrollo en Criotecnología de Alimentos (CIDCA e CCT La Plata
e CONICET) e Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP),
Buenos Aires, Argentina
Irene Domínguez
M.J. Fabra
Preservation and Food Safety Technologies Department, Institute of Agrochemistry and Food
Technology (IATA-CSIC), Valencia, Spain
Rafael Gavara
Laura G. G
omez-Mascaraque
Food Chemistry and Technology Department, Teagasc Food Research Centre, Moorepark,
Ireland
Stefania Gottardo
xiii
xiv Contributors
Pilar Hernández-Muñoz
K.C. Ho
Department of Chemical and Process Engineering, Faculty of Engineering and Built
Environment, Universiti Kebangsaan Malaysia, Bangi, Malaysia
Eddo Hoekstra
Taylor Ingle
National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR,
United States
Seid Mahdi Jafari
Department of Food Materials and Process Design Engineering, Gorgan University of
Agricultural Sciences and Natural Resources, Gorgan, Iran
Ali Kermanizadeh
Heriot Watt University, School of Engineering and Physical Sciences, Nano Safety Research
Group, Edinburgh, United Kingdom; University of Copenhagen, Department of Public Health,
Section of Environmental Health, Copenhagen, Denmark
Jesper T.N. Knijnenburg
International College, Khon Kaen University, Khon Kaen, Thailand
Katrin Loeschner
Technical University of Denmark, National Food Institute, Division of Food Technology,
Lyngby, Denmark
Amparo L opez Rubio
Gracia López-Carballo
Olga Martín-Belloso
M. Martínez Sanz
Adriana N. Mauri
A.W. Mohammad
Environment, Universiti Kebangsaan Malaysia, Bangi, Malaysia; Research Center for Sustainable
Process Technology (CESPRO), Faculty of Engineering and Built Environment, Universiti
Kebangsaan Malaysia, Bangi, Malaysia
Contributors xv
Peter Møller
University of Copenhagen, Department of Public Health, Section of Environmental Health,
Copenhagen, Denmark
Thilak Mudalige
AR, United States
Yanina S. Musso
Suresh Neethirajan
BioNano Laboratory, School of Engineering, University of Guelph, Guelph, ON, Canada
Isabel Odriozola-Serrano
Gemma Oms-Oliu
Angel Paredes
National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR,
United States
Ruud Peters
RIKILT, Wageningen UR, Wageningen, The Netherlands
Lidija Posavec
Previously: Department of Health Sciences and Technology, Swiss Federal Institute of
Technology Zurich, Zurich, Switzerland
Haiou Qu
AR, United States
K.V. Ragavan
BioNano Laboratory, School of Engineering, University of Guelph, Guelph, ON, Canada
K.S.M.S. Raghavarao
Academy of Scientific and Innovative Research, CSIR-CFTRI, Mysore, India; Department
of Food Engineering, CSIR-Central Food Technological Research Institute (CFTRI),
Mysore, India
Kirsten Rasmussen
Hubert Rauscher
N.A. Rosnan
Environment, Universiti Kebangsaan Malaysia, Bangi, Malaysia
xvi Contributors
Pablo R. Salgado
on y Desarrollo en Criotecnología de Alimentos (CIDCA e CCT La
Plata e CONICET) e Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP),
Reinhilde Schoonjans
European Food Safety Authority, Parma, Italy
Richa Sharma
Academy of Scientific and Innovative Research, CSIR-CFTRI, Mysore, India; Department of
Food Engineering, CSIR-Central Food Technological Research Institute (CFTRI), Mysore,
India; Department of Biotechnology, Sharda University, Greater Noida, India
Alexandra Teleki
Department of Pharmacy, Uppsala University, Uppsala, Sweden
Y.H. Teow
Environment, Universiti Kebangsaan Malaysia, Bangi, Malaysia; Research Center for Sustainable
Process Technology (CESPRO), Faculty of Engineering and Built Environment, Universiti
Kebangsaan Malaysia, Bangi, Malaysia
Desiree Van Haute
AR, United States
Eveline Verleysen
Service Trace Elements and Nanomaterials, Sciensano, Brussels, Belgium
PREFACE
The last decades have witnessed an increased interest in the use of nanomaterials in a
number of different scientific and industrial areas, including the food industry. In fact, the
use of nanomaterials for food applications is a rapidly evolving field and, given the specific
properties of nanomaterials and their tremendous potential, an increased number of ma-
terial innovations which contribute to improved food quality and safety are foreseen.
In this book, we have tried to compile the latest developments reviewed by
renowned experts related to the various uses of nanomaterials in the food area, to set
the basis for future inspiration. Although the possibilities are unlimited, nanomaterials
in the food area could be divided in several broad groups, corresponding to different sec-
tions within this publication. The first group consists of nanomaterials for improved food
quality. In this sense, the use of nanoingredients to enhance nutrient absorption or nano-
technology tools (like nanoemulsions or nanoencapsulation) for functional food develop-
ment has proved successful. The challenges in this area rely on a more thorough
knowledge for the proper selection of materials to attain the desired properties in terms
of protection, release, and digestion. In vivo behavior of these nanoingredients within
complex food matrices is another area which deserves investigation, as a few works
have demonstrated that the properties of the nanomaterials can be modified as a conse-
quence of their interaction with other food components. The second big group relates to
nanomaterials for improved food safety. Nanosensors and nanocomposites for food
packaging applications are some of the nanomaterials which can be included in this
group. Regarding the nanosensors, the ability of nanoparticles to be functionalized,
together with their recognition and transducing properties, and the possibility to
combine them with biosensors opens up a great number of possibilities to develop highly
sensitive tools that guarantee food safety (there are a couple of chapters devoted to these
interesting issues). In the food packaging area, specifically for polymeric and bio-
polymeric materials, incorporation of well-dispersed nanoparticles (organic and inor-
ganic) has demonstrated their ability to improve mechanical, thermal, and barrier
properties without affecting the optical characteristics of the materials. This is probably
the area within the food nanotechnology field where more research has been done giving
rise to various commercial products. From several years ago, there have been research ef-
forts put on going a step further and, thus, the use of these nanoparticles are aimed, not
only at improving the quality and safety of the packaged food passively, but at playing an
active role in food preservation and food quality enhancement. These novel materials are
known as active/bioactive packages, which are able to release/absorb certain substances
to change either the inner packaging atmosphere or even the food product. The aim of
xvii
xviii Preface
combining these active/bioactive substances included in the packaging structure with

nanoparticles is to modulate their release or sorption or even to use the nanoparticles
as active substances themselves (such as antimicrobial packages containing nanometals).
However, it is also true that very little is known about the potential migration of nano-
particles from the packages and their subsequent potential toxicity, and there is also con-
troversy regarding the use of nanoingredients and the need of “nanolabelling” the food
products. Therefore, a complete section written by toxicology experts with clues about
the toxicological assessment of food-related materials containing nanoparticles and legis-
lation authorities (both from Europe and the United States) has been included in the
book.
We sincerely hope that you enjoy the book as much as we have enjoyed editing it.
The Editors
Valencia
CHAPTER 1
Nanomaterials for Food Applications:

General Introduction and Overview of
the Book
Amparo Lo pez Rubio1, Laura G. Go
mez-Mascaraque2, M.J. Fabra1,
1
M. Martínez Sanz
1
Preservation and Food Safety Technologies Department, Institute of Agrochemistry and Food Technology (IATA-CSIC),
Valencia, Spain; 2Food Chemistry and Technology Department, Teagasc Food Research Centre, Moorepark, Ireland
Contents
1.1 Introduction 1
1.2 Potential of Nanotechnologies to Improve Consumer Health 3
1.3 Nanotechnologies for Food Processing and Sensing Applications 4
1.4 Food Packaging Nanotechnologies 6
1.5 Characterization, Toxicological Assessment, and Regulatory Status of Nanomaterials in Food 7
References 8
1.1 INTRODUCTION
Nanotechnology refers to the manufacturing, characterization, and manipulation of ma-
terials with at least one of the dimensions in the nanometer scale (below 0.1 mm).
Although the description of nanotechnology is purely in terms of the size, it is important
to emphasize that, at this scale, rearranging the atoms and molecules leads to changes in
physicochemical properties in comparison to the same material at larger size scales. Spe-
cifically, the decrease in size has associated much larger specific surface area with a sub-
sequent enhanced surface reactivity and, for instance, increased ion release (Peters et al.,
2016). Applications of nanotechnologies in the food sector are rapidly growing, with an
amazing boost on the number of the publications, patents and intellectual property rights
in the field, and interesting nanotechnology-related developments in food processing,
packaging, nutraceutical delivery, quality control, and functional foods among others
(Dasgupta et al., 2015). In fact, nanotechnology is considered to be one of the six key
enabling technologies for Europe (European Commission, 2016), given its potential
for the development of innovative products and applications in several industrial sectors
including food production, food processing, novel foods, food additives, and food con-
tact materials (Chaudhry et al., 2008; Kah et al., 2013; Kah and Hofmann, 2014; Sekhon,
2010, 2014).
Nanomaterials for Food Applications © 2019 Elsevier Inc.

ISBN 978-0-12-814130-4, https://doi.org/10.1016/B978-0-12-814130-4.00001-4 All rights reserved. 1
2 Nanomaterials for Food Applications
Investment in nanotechnology has burst in the past years. In fact, nearly every major
funding agency has a budget devoted to this research area, and it is expected that by 2020,
products incorporating nanotechnologies will contribute approximately $1 trillion to the
global economy, being about two million workers employed in nanotech industries, and
three times that many having supporting jobs.
The two existing approaches to generate nanoparticles or nanodevices are the top-
down approach (i.e., breaking up bigger particles) or the bottom-up approach that con-
sists of atom-by-atom engineering. This last strategy can be accomplished either using
nanoscale engineering tools (such as electron-beam and ion-beam fabrication or nanoim-
print lithography) or through the self-assembly of molecules such as in the case of amphi-
philic molecules to form micelles. And regarding the characteristics that make
nanomaterials so attractive, the first one is, of course, related with the size, as such small
sizes have advantages both from economical point of view and application-wise. How-
ever, as mentioned before, at these mesoscales new phenomena occur, which is relevant
in areas such as electronics. Moreover, the ability of arranging atom-by-atom provides
with an extremely high design control, allowing for huge possibilities in terms of product
development.
In the food area, nanotechnologies can be applied along the whole cycle of product
development, from the production of nanoingredients or nanofertilizers for food produc-
tion, through the use of nanotechnologies for food processing (for instance, nanofiltra-
tion technologies) to even the use of nanosensors or nano-noses to assure food quality
and safety.
According to several consulting agency reports, in the food area, the overall size of the
global nanotechnology market is estimated to be worth more than $20 billion with an
upward trend and being led by the United States, followed by Japan and China
(Cientifica Report, 2006; Helmut Kaiser Consultancy, 2004; Nanoposts Report,
2008). Most of the food giants including Nestle, Kraft, Heinz, and Unilever support spe-
cific research programs to capture a share of the nanofood market in the next decade.
The main food-related areas where nanotechnologies have a great potential are food
packaging (nanocomposites, active, bioactive, and intelligent packaging), functional
foods, and nutraceuticals (through the use of nanoencapsulation or nanoformulations),
food processing (using technologies such as nanofiltration), and food safety and quality
(through the use of nanosensors, nanotongues, and nano-noses). However, for the suc-
cessful establishment of nanotechnologies in the food sector, a number of technological,
societal, and regulatory barriers need to be overcome. The real benefits of these technol-
ogies for consumers and the evaluation of the potential risks associated to consumption
of, or exposure to nanoparticles need to be deeply analyzed and communicated to
society.
Therefore, this book was conceived to provide current knowledge about the benefits
that these novel technologies may have in the food sector, as well as their current
Nanomaterials for Food Applications: General Introduction and Overview of the Book 3
regulatory status and the challenges encountered when trying to characterize and/or
quantify the nanomaterials present in complex food matrices.
1.2 POTENTIAL OF NANOTECHNOLOGIES TO IMPROVE CONSUMER

HEALTH
One of the areas in which nanotechnologies play a relevant role in the food sector is the
development of functional foods, which are food products especially designed to provide
health benefits beyond their basic nutritional value (Day et al., 2009). The design of these
products usually involves the addition of exogenous bioactive ingredients to a conven-
tional food matrix (Kaur and Das, 2011), and this entails a series of challenges. First of
all, most bioactive ingredients are sensitive and can lose activity during manufacturing,
storage, commercialization, and/or consumption (G omez-Mascaraque et al., 2015).
Moreover, some of these compounds are immiscible with the food matrix in which
they are to be incorporated, some can alter the organoleptic properties of the final
food products, and many of them have poor bioavailabilities (Deng et al., 2014; Lafarga
and Hayes, 2017). These limitations can only be overcome through adequate formula-
tion and/or encapsulation strategies to protect, transport, and deliver the ingredients of
interest. In this context, nanotechnologies are a promising alternative to conventional
techniques because the miniaturization of the delivery vehicles generally results in a
reduced impact on the textural properties of the products, an improved product stability,
and an enhanced absorption of the bioactive ingredients, due to their greater surface-to-
volume ratio (McClements, 2015).
There are already several commercial food products in the market containing nano-
sized delivery vehicles, such as the “Tip-Top” Up bread, commercialized in Australia,
which contains fish oil nanocapsules, which are a source of omega-3 fatty acids. The cap-
sules are designed to break open only when they have reached the stomach, thus avoiding
the unpleasant/strong taste of fish oil. Another commercial example is from the Israeli
Company NutraLease, which uses expanded micelles (hollow spheres made from fats,
with an aqueous interior) with a diameter of approximately 30 nm. The nutrients or
“nutraceuticals” are contained within the aqueous interior. The technology has already
been adopted and marketed by Shemen Industries to deliver Canola Active oil, which it
claims reduces cholesterol intake into the body by 14%, by competing for bile
solubilization.
An example of nanotechnologies used in the nutraceutical sector is the Spray For Life
product range manufactured by Health Plus International. They use a newly designed
Nanoceutical Delivery System for oral administration of dietary supplements, resulting
in increased bioavailability compared with gastrointestinal absorption.
Apart from the aforementioned commercial food products, a number of nanosized
delivery vehicles and formulations have already been developed and are currently being
investigated for their potential use for food applications. These include nanoemulsions,
nanoliposomes, lipid nanoparticles, biopolimeric nanoparticles and coacervates, and
nanohydrogels. Chapter 2 of the present book provides an overview of the different
nanostructured delivery systems, which can be applied for the formulation of food ingre-
dients, with a special emphasis on the impact that their nanostructure and physicochem-
ical properties have on the bioavailability of the functional ingredients entrapped within
them. In general, a reduction in the droplet or particle size increases their bioaccessibility
(Yallapu et al., 2011). A review of recent research works evaluating the role of nanofor-
mulation and nanoencapsulation of bioactive food ingredients on their bioaccesibility and
bioavailability is also provided in the second chapter, summarizing findings obtained from
experiments conducted both in vitro and in vivo. Finally, their industrial applications are
also discussed.
The classification of the nanoencapsulation techniques used to produce the diverse
range of nanostructured delivery vehicles commented above is complex and variable
because of the diversity of existing technologies. Nevertheless, Chapter 3 aims at
providing a systematic classification and description of these technologies by dividing
them into five different groups ( Jafari, 2017). These include nanoencapsulation within
biopolymer nanoparticles, lipid-based nanoencapsulation systems, nature-inspired nano-
carriers (such as caseins, cyclodextrins, or amylose nanohelices), novel nanoencapsulation
systems produced using advanced equipment and technologies (such as electrospinning,
electrospraying, nanospray-drying, or micro- and nanofluidics), and other miscellaneous
nanoencapsulation systems such as nanocrystals, dendrimers, or niosomes. The applica-
tion of these technologies for the nanoencapsulation of different bioactive ingredients,
including phenolic compounds, antimicrobial agents, bioactive oils, carotenoids, and
other food colorants, among others, is also reviewed in the third chapter.
Given that minerals and vitamins are essential nutrients for humans and that insuffi-
cient intake of these micronutrients affects billions of people worldwide (Bailey et al.,
2015), Chapter 4 of this book is especially dedicated to the nanoformulation of these
bioactive ingredients for food fortification and supplementation purposes. Again, by
decreasing the particle size down to the nanoscale, the bioavailability of minerals and vi-
tamins can be improved, although the impact of nanostructuring these ingredients on
their toxicity must be also evaluated because it may result in adverse effects.
1.3 NANOTECHNOLOGIES FOR FOOD PROCESSING AND SENSING

APPLICATIONS
Nanotechnology has also brought significant advances within the food processing area.
Because of the high-quality standards required for food products and the increasing de-
mand on emerging products such as low fat and low calorie products and foodstuffs
adapted for special dietary requirements, improved separation methods are needed within
the food industry. One of the most relevant nanotechnology-based methods with a great
potential for food processing applications is nanofiltration. Membranes with nanopores
can be used, for instance, for water purification and softening, as such nanofilters are
able to remove divalent ions. This nanotechnology is also very useful in the dairy industry
for several applications, such as to fractionate milk proteins, for enhancing the microbial
quality of dairy fluids (as the membranes also remove bacteria), and for the standardization
of milk. Among the several advantages of this technique over conventional filtration pro-
cesses, it is worth highlighting its lower energy consumption, lesser processing steps,
greater separation efficiency, and improved final product quality. Chapter 5 presents
the basic principles of the nanofiltration process, followed by an overview of the recent
developments of this processing method for several industrial applications such as dairy
products, beverages, sugar, wastewater treatment, and vegetable oils, as well as the chal-
lenges that need to be addressed to improve nanofiltration for its large-scale application
within the food industry.
Nanotechnology has also led to significant advances within the food safety area.
Currently, there is a high demand on robust, fast, and sensitive detection techniques
for food contaminants. In this context, biosensing techniques, which rely on highly spe-
cific bioreceptors coupled with efficient transducing systems, are the most efficient
approach for the specific recognition of food contaminants, in the presence of highly
complex food matrices. These bioreceptors need to be strongly and selectively binding
biomolecules for a specific target compound, producing a chemical or biological signal
that is converted to a measurable form by the transducer. Aptamers are single-stranded
nucleic acid or peptide molecules that have high affinity to only their target ligands,
similar to antibodies, but much simpler to synthesize and modify. One commercial
example of biosensing for the detection of food contaminants is the Nanobiolumines-
cence Detection Spray from the company Agromicron, which is based on a luminescent
protein that emits a visible glow when it reacts with the surface of some pathogenic mi-
croorganisms such as Salmonella or Escherichia coli.
Chapter 7 presents an overview of the unique physicochemical advantages of nano-
materials as efficient signal transducers and amplifiers, in conjunction with the selectivity
and flexibility of aptamers. The different nanomaterial-based aptasensing techniques for
food safety are presented and some of the most relevant examples of application are also
discussed in this chapter.
Food quality and safety are the two most important aspects focused in food processing
industry. Analytical procedures and instrumentation are sufficiently equipped to deter-
mine food quality and hazards (contaminants and adulterants) at very low concentration
with great accuracy in complex food matrices. However, because these instruments and
their services are expensive, they required trained/skilled personnel and they are time-
consuming to obtain the results. Hence, there is a need for simple, affordable, field appli-
cable sensor systems with sufficient analytical performance for the evaluation of food
safety and quality. Chapter 8 reviews the interest of biosensors for fulfilling the above re-
quirements and with the advent of nanotechnology. The discovery of nanomaterial
properties in recent times and their integration with biosensors have further helped to
progress in the field. This chapter presents an overview of various components of biosen-
sors, their properties, and recent advances with respect to food safety and quality. At the
end, perspective and scope of nanomaterial properties along with challenges in the field
of biosensors for food industry are also discussed.
1.4 FOOD PACKAGING NANOTECHNOLOGIES

Within the food-packaging sector, the nanocomposites area is the most active one in
terms of food nanoscience research. The use of biodegradable plastics and resources
are seen as one of the many strategies to minimize the environmental impact of
petroleum-based plastics. However, most of the biobased packaging materials do not
perform as the petroleum-based ones and some of their physical properties such as barrier,
thermal stability, and mechanical properties need to be improved. The nanoreinforce-
ment of biobased plastics through the incorporation of nanoclays, micro- and nanofibers
of cellulose, and carbon nanofibers and nanotubes has been proven to be an effective way
to enhance these properties concurrently (Sanchez-García et al., 2010; Trifol et al., 2016;
Vilarinho et al., 2017). Some of the advantages of using nanofillers instead of the previ-
ously used microfillers is that with very low percentages of loading, typically below 10%,
relevant physical improvements can be achieved without compromising desirable matrix
properties such as transparency or toughness (Mondragon et al., 2015; Vilarinho et al.,
2017). Moreover, the greatest specific surface of the nanofillers improves the adhesion
with the matrix and thus better final properties can be obtained. All these aspects,
together with the environmental impact of nanotechnology applied to food packaging
are discussed in more detail in Chapter 9. Moreover, the application of metallic nanopar-
ticles with antimicrobial or antioxidant properties embedded in polymeric films for the
development of active packages is also addressed in this chapter.
New technologies of food packaging try to respond to society demands. Thus, bioac-
tive packaging has the unique role of enhancing food impact over consumers’ health
through the generation of healthier packaged foods. In this sense, it can be considered
as an innovative strategy in the production of functional foods. The use of nanotechnol-
ogies also opened new possibilities for improving the effectiveness of these packages, and
many nanoparticles have been developed because of their potential to encapsulate active
compounds and increase their functionality, stability, and bioavailability. The incorpora-
tion of nanoparticles within the packaging structure, together with the active or bioactive
substances, would permit a controlled release of these additives, diminishing the impact
on materials properties (Imran et al., 2010; L
opez-Rubio et al., 2006). Chapter 10 focuses
on the application of nanotechnology in the development of bioactive packaging. It
analyses the requirements of current food packaging, describes the main bioactive
compounds for the preparation of bioactive packaging and studies the development of
nano-sized carrier systems and different strategies in the preparation of nanostructured
packaging materials to improve the stability, dispersibility, availability, and transport of
bioactive compounds.
1.5 CHARACTERIZATION, TOXICOLOGICAL ASSESSMENT, AND

REGULATORY STATUS OF NANOMATERIALS IN FOOD
To understand the challenges for proper characterization and risk assessment of nano-
technologies in food, it is important to emphasize that nanoparticles can be either natu-
rally present in the food products, intentionally added, nonintentionally incorporated
(either through migration or environmental contamination), or can even be formed dur-
ing food processing. A clear example of a naturally containing food is milk, where casein
micelles of about 100 nm coexist with whey proteins having a size of about 30 nm. In
fact, many food proteins exist naturally in the nanoscale and simple triglyceride lipids
are about 2-nm long. We can also find nanoparticles in traditionally processed food prod-
ucts, especially in emulsions such as mayonnaise. Furthermore, during digestion, food is
broken down to nanostructures before assimilation and, thus, the food compounds are
metabolized in the body at a nanoscale. And although it is still unclear whether nanoscale
processing of food materials might produce structures that are different from those that
occur naturally, a fair classification before evaluating potential risks should be carried
out to clearly distinguish between nanoparticles produced from food-grade materials
and inorganic or engineered nanoparticles (such as nanometals or carbon nanotubes
and nanofibers). Anyway, considering not only the different potential sources of nano-
particles and the inherent complexity of food matrices, for nanoparticle quantification
and characterization, a proper sample preparation has to be carried out and the selection
of the analytical tools are key for obtaining reliable results. All these aspects are dealt with
in Chapters 10 and 11.
Regarding the factors related to toxicity, without any doubt the dose is the main fac-
tor to take into account. Other important factors are the entrance route and certain
intrinsic factors that depend on the type of nanoparticle, such as the size, shape, compo-
sition, charge, and if nanoparticles are coated, aggregation, biodegradability, bio-
distribution, etc. All these aspects, together with the current knowledge about the
health impact that certain nanoparticles may have on ingestion or exposure are discussed
in more detail in Chapter 12.
There are several international agencies that have elaborated protocols or guides for
nanotechnology-related risk evaluation, such as the French OECD or the International
Standards Organization. The European Food Safety Authority published a guide for risk
analysis in food products, which include the identification and characterization of
nanomaterials and the identification of risks, both in vitro and in vivo. This guide (see
Chapter 13) includes a series of tests that should be carried out, some of them necessary
and others specific for certain applications. Although the number of research works about
nanoparticle toxicological assays has increased considerably in the last years, experts in the
area have found that many of them have been carried out by nonexperienced researchers,
being incomplete, using excessive nanoparticle doses or leading to wrong conclusions.
On the other hand, food industries are still reticent to publically declare if they work
or use nanotechnologies (probably due to potential consumer rejection) and, thus, to
ensure the proper spreading of these nanotechnologies, a standardization of the method-
ologies for risk and toxicity assessment to guarantee the safety of any material to be
included in food products deems necessary.
The details about the current regulatory status of nanotechnologies in the European
Union are compiled in Chapter 13.
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need expect nothing to come of it. To-day I am in doubt.... I desired
to hold myself free from—er—such things as marriage. Doubtless
that, also, was a mistake.... I am open-minded.”
“You—you—are open-minded!” Carmel gasped out the words.
“Exactly. I have determined to allow the emotion to follow its natural
course, without interference by myself. Even if it results in marriage
with you, I shall not interfere.”
“Of all things,” said Carmel.
“Meantime, while the more important matter is working itself out, let
us endeavor to be friends.” As he said this there came into his voice
a wistfulness, a humility which touched her. Her eyes filled. She held
out her hand.
“Friends!... Of course we shall be friends! You must overlook my bad
temper. I have so many faults.”
His eyes glowed, his face became animated. “You,” he said, eagerly,
“are very lovely. You are—er—wonderful....” He stared at her as if
she had been an apparition. Carmel caught her breath and turned
away abruptly.
So much for Evan Pell’s effort to break through his chrysalis shell....
The fates had not determined if he were to become a moth or a
butterfly....
At that very hour Abner Fownes was opening his mail. His frame of
mind was not of the pleasantest, though he had succeeded in tiding
over the day before a situation financially threatening. The condition
of his affairs was wearing upon him. Constant calls for money,
demands upon his shiftiness to prevent a débâcle, never-failing
watchfulness, bore heavily upon the man. It was not easy to maintain
his attitude of high-spirited public citizen. It was not simple to keep
beneath the surface the man who lurked under the skin of the
fatuous cat’s-paw. It was difficult to maintain the pretense of being
used by smaller men, when constantly he had to twist smaller men to
his own ends.
Now he opened with trepidation a letter from a lumber concern with
which his dealings had been extensive.
We have received yours of the 20th with respect to
renewing your note for $18,750 which falls due two weeks
from to-day. We regret that in present conditions this is
impossible, and must ask you to take up this paper without
fail.
Fownes crumpled the letter in his hand and stared at the paneling of
his office as if he hoped by the mere venom of his look to reduce it to
ashes. His pudgy, beautifully tailored shoulders moved upward so
that his short neck disappeared and his ears rested upon his collar.
Then he expelled his breath. He arose and went to the safe, which
he opened—to which he alone possessed the combination—and
took from its resting place the red leather book in which he kept the
true record of his and his company’s condition. This he carried to his
desk, and for many minutes he studied it, hoping against hope for
some expedient to make itself apparent.... There was no expedient.
He returned the book to its place and locked the safe; then he
twisted the handle of the telephone insistently, and gave Central the
number of the Court House.
“Deputy Jenney,” he said, arrogantly.
The deputy answered.
“Come to my office immediately,” he said. “Never mind who sees.
This is imperative.... At once.” Following that, he waited.
Deputy Jenney entered, breathless, and stood panting.
“Jenney,” said Fownes, “I’ve determined to make another
investment.”
“Eh? Already.... Why, we hain’t hardly got the last off’n our hands. It’s
takin’ a chance, says I, and crowdin’ the mourners.”
“I’m running this business, Jenney.... This next is to be no retail deal,
either. It’s wholesale.”
“You—you want to go easy. By golly! Mr. Fownes, so much stuff
comin’ in is goin’ to git somebody mighty curious.”
“If you’re sheriff, Jenney, what will the curiosity amount to?”
“Federal officers!”
Fownes shrugged his shoulders. “I’ll see to your appointment as
sheriff. You attend to organizing everybody to receive the shipment.
We’ll need all available space and all outlets. I’m going to fetch in
enough this time to flood the county.”
“You know what you’re doin’,” Jenney said, sullenly, “but what with
that damn paper a-peckin’ at us all the time——”
“Nobody reads it, Jenney. And you’ll be sheriff.”
“I’ll do my dumdest—but I don’t like it.”
“And I don’t care whether you like it or not. And that’s that. Better see
Peewee first.”
“When’s it comin’?”
“Inside of ten days.... And, Jenney, I don’t believe the paper’s going
to bother much longer.”
“Eh?”
“I’m going to—er—give that girl a hint of our plans.”
“What?”
“I’m going to give her a tip, as they say. She’ll investigate, and that
professor will investigate.”
“Like Sheriff Churchill did?”
“The result,” said Fownes, “will be similar.”
CHAPTER XVI
“YOU are not,” said Evan Pell to Carmel Lee, “familiar with
laboratory practice—er—with chemical analysis, for instance.”
“I know nothing about it?”
“I judged not,” he said, unwittingly reverting to his patronizing
manner. “However, it seems to me the individual who searches for
truth—in the happenings of the day—would be better fortified for his
labors if he applied the methods of the chemist.”
“As, for instance?”
“Let us suppose there has been a crime. The crime is a result. An
inevitable result of the combination of certain elements. Given the
crime, the chemist should be able to analyze it and to separate its
elements.”
“I believe that is the method of story-book detectives.”
“No.... No.... This is science, logic. A simple example. You hold a
substance in your hand. You moisten it with iodine. If the substance
turns purple you know starch is present. Do you see?”
“I’m sure I don’t see.”
“What do you think of Abner Fownes?” he asked, with
uncharacteristic swerving from the subject.
“I think he is abominable.”
“Possibly.... But impersonally, as an individual—what then?”
“He is a pompous, self-deceiving, hypocritical poseur.”
“Uh!... As to intelligence?”
“As your chemist would say—a trace.”
“I fear,” he said, “you have neglected to moisten him with iodine.”
“What do you mean?”
“I mean,” he said, “that you have utterly failed to comprehend what
you are facing—what it is you have to do in this headstrong crusade
of yours against the liquor smugglers.”
“What has that to do with Abner Fownes?”
“That,” he said, “is the big question.”
“But why should he? That is absurd. Perhaps the smugglers are
using him as a cat’s-paw in some manner—but he’s rich. There’s no
need. These men take the risks they must for profit.”
“Miss Lee,” he said, “you—er—challenged me to investigate this
affair. I promised to do so.... I have set about it in an orderly manner.”
“So I imagine,” she said, a trifle wearily.
“I have started with the compound itself—with the fact that we know
there exists a wholesale traffic in liquor, from which a huge profit is
derived. This is compounded of many small elements. I think we may
take it as fact that the hunchback, Peewee Bangs, is an element;
that his hotel is another element; that Deputy Jenney is a rather
important ingredient. For myself, I am satisfied numerous citizens of
Gibeon are involved—in the distribution and marketing of the liquor. I
am quite certain, for instance, that the business of taking
photographs is not the sole means of livelihood followed by Lancelot
Bangs.... He is, I believe, a cousin to the proprietor of the Lakeside
Hotel.... These things are present in the compound, but they could
not, joined together, cause the result we see. The principal ingredient
is missing.”
“And what is that?”
“A daring, ruthless intelligence. Able leadership. The brain capable of
conceiving of bootlegging as an industry, and not as a matter of petty
retailing.”
Carmel Lee was impressed. Evan Pell possessed the quality of
holding interest, of seeming to speak from sure knowledge.
“I think you are right so far. What we need is to find this intelligence.”
“I rather fancy I have found him. In fact, I have had little doubt as to
his identity for a considerable time.”
“Abner Fownes?” She shrugged her shoulders. “I dislike him—he is
insufferable—but the idea is absurd. Bumptious little men like him,
secure in their wealth and position, do not jeopardize it.”
“That,” said Evan, “is dependent upon their security. What would you
say if I were to tell you Abner Fownes has been on the brink of
bankruptcy for months? What would you say if I told you this rum
running commenced only after his finances became tangled? What
would you say if I told you the major part of the profits from this liquor
business went to maintain Abner Fownes in the character he has
assumed, and keep his imperiled business out of the hands of his
creditors?”
“I would say,” she said, “that you are crazy.”
“Nevertheless,” he said, “I am convinced of the fact.”
“But he has no brains. Look at him. Observe him.”
“Miss Lee, it takes a man of tremendous resolution and of very keen
intelligence to invent for himself a character such as he has exhibited
to Gibeon for years.... If the world supposes you are a nincompoop—
a vain figurehead—a puppet set up by other men—you are little in
danger of arousing suspicion as to yourself. When a man is
commonly admitted to be a fool, he is safe. Fownes has been at
infinite pains to prove himself a fool.”
Carmel was far from dull. Her mind flashed to the keystone of the
arch Pell was constructing. “Show me he is on the verge of
bankruptcy and maybe you can convince me of the rest.”
He told her. He itemized the contracts Fownes had made for the
purchase of lumber, and the prices at the time of sale. He showed
how the market had declined, and the total sum of Fownes’s losses.
“These,” he said, “are facts—not public, but easy to come by.... I first
found the trail of them when the cashier of the bank asked me to
assist him in an audit of the books. That was some months ago
when I occupied my official position.”
“But if you are right, then Abner Fownes is a murderer, or an
instigator of murder.... Nobody can look at him and credit that.”
“Abner Fownes,” said Pell, “is capable of any crime to preserve
Abner Fownes. I have watched him, studied him. I know.”
“I can’t believe.... It is incredible. No. You must be mistaken.”
“Miss Lee,” said Pell, solemnly, “if you wish to continue to exist, if
you hope to come through this affair with your bare life, you must
believe. If you cannot believe, pretend it is a fact and act accordingly.
Forget everything else and concentrate upon Abner Fownes.... But
take this warning: The moment he suspects you suspect him—you
will doubtless join Sheriff Churchill.... I believe Churchill was on the
road to the discovery. He would not have disappeared otherwise.”
Carmel remained silent, considering. At length she spoke. “You are
right,” she said. “One does not insure his house because he believes
it will burn, but in case it shall burn. I shall make believe you are right
about Abner Fownes—as an insurance policy.... But where does that
lead us?”
“To the sheriff’s office,” said Pell.
“What?”
“If Jenney is appointed sheriff to succeed Churchill, where is the
machinery to fight Fownes? He could laugh at us. Therefore Jenney
must not be appointed.”
“But how can that be averted?”
“I think,” he said, “the sole hope lies in yourself.”
“In me!”
“You must find a man, a man of courage, of public spirit. You must
find a man who can be relied upon and whose name will carry weight
with the Governor.... When you find him, you must go to the Capitol
and make the Governor appoint him—and you must act at once.”
“I?... I go to the Governor?”
“You.... If you could carry a petition, signed by a number of citizens, it
would strengthen you, but I don’t see how that can be done.... And
yet—and yet——”
“It must be done.... Secretly.”
“To approach one man—who would talk, who was on the other side
—would be to ruin the whole project.”
“Nevertheless, it must be done.”
He shrugged his shoulders. “First find a man worthy to hold the
office,” he said.
“I shall find him.... I know Mrs. Churchill. She will know her
husband’s friends and supporters—the men who worked for his
election and whom he trusted.”
“The idea is good,” said Evan. “Suppose you act without delay.”
Carmel found Mrs. Churchill in the kitchen, giving a hearty welcome,
in spite of her baking, to the visitor.
“Set,” said Mrs. Churchill, “and lemme pour you a cup of tea. Always
keep it simmerin’ on the back of the stove in case of headache.” This
was a favorite fiction of Mrs. Churchill’s—that she suffered with her
head and that tea was the only remedy. It would appear, however,
that she used the beverage as a preventive instead of a cure.
“I’m sorry, but I haven’t time to sit this morning. I’ve come to you
because you’re the only person in Gibeon who can help me—and
because you are the one most interested in helping me.... I want to
know whom to trust.”
“Eh?... Trust? Speakin’ of young men, be ye?”
“No.” Carmel smiled as she saw the fire of matchmaking light Mrs.
Churchill’s motherly eyes. “I want to know whom your husband
trusted. I’ve got to find a man.... Deputy Jenney is going to be
appointed sheriff,” she said.
Mrs. Churchill’s eyes flashed. “In my man’s place! That critter!”
“If,” said Carmel, “I can’t find an honest man—one like your husband
—and get there first.”
“Uh huh....” said Mrs. Churchill, ruminatively. “He wa’n’t much give’ to
talkin’, but more’n once he says to me, says he, ‘The’ hain’t many in
this place I’d trust as fur’s I could throw ’em by the horns,’ he says.
But I call to mind that whenever he got kind of out of his depth like,
and had to talk things over with somebody, he allus went to spend
the evenin’ with Jared Whitefield. Him and Jared was close. I don’t
calc’late you’d make no mistake goin’ to Jared and statin’ your case.”
“Thank you,” said Carmel. “There’s not a moment to be wasted.
Good-by.”
She did not return to the office, but walked out the main street, past
the village cemetery, to the rambling, winged house from which
Jared Whitefield ruled his thousand-acre stock farm—a farm he had
carved himself out of the forest, cleared, stumped, and planted. She
knew the man by sight, but had never held conversation with him. He
was not an individual to her, but a name. She opened the gate with
trepidation, not because she feared the reception of herself, but
because she was apprehensive. Mr. Whitefield, when studied at
close range, would not measure up to the stature of the man she felt
was needed.
A dog barked. A voice silenced the dog. Carmel noted how suddenly
the dog quieted when the voice spoke. Then a man appeared
around the corner of the house, an ax in his hand, and stood
regarding her. He did not bow, nor did he speak. He merely stood,
immobile, as if some cataclysm of nature had caused him to burst
through the soil at that spot, and as if there still remained embedded
roots of him which anchored him forever to the spot. He was big,
straight, bearded. At first glance she thought him grim, but instantly
decided it was not grimness, but granite immobility. She approached
and greeted him.
“Good morning, Mr. Whitefield,” she said.
He inclined his head and waited.
“I am Miss Lee, proprietor of the Free Press,” she said.
“I know ye,” he said.
Surely he was difficult; but for all that, she felt herself drawn to the
man. There was a feeling that if she could scale his granite sides
and sit upon the shelf of his shoulder she would be safe—that
nothing could topple him from the spot where he had taken root.
“I want to talk to you, Mr. Whitefield. It is a matter of great importance
—almost of life and death,” she said.
“Say it,” said Jared Whitefield.
“They’re going to appoint Deputy Jenney sheriff,” she said.
“Know it.”
“It mustn’t be.”
“Why?”
“Is it safe to speak here. A word overhead——”
“This is my yard,” said Jared, and there was much, much more in the
words than the mere statement of the fact. It was a declaration of
independence. It was a guaranty. It lifted Jared out of the
commonplace and made a personage of him—the unquestioned
ruler of a principality. Where he was, he ruled.
“You know what my paper has done.”
“Lighted matches nigh a powder keg.”
“I believe, and I hope to prove, there is an organization here for the
purpose of wholesale dealing in smuggled liquor. I believe that
organization murdered Sheriff Churchill. I believe Jenney is a part of
it and that his appointment as sheriff is a move to give the criminals
safety in their work. I know there are huge profits. At the top is some
man of intelligence who directs. I want to get that man.”
“Who?”
“I think Sheriff Churchill knew—or guessed. That’s why he is dead.”
“Uh!... Wa-al?”
“Our only chance is to block Jenney’s appointment. To get first to the
Governor with the name of another man—a man whose name and
personality carry weight. If we can get the office of sheriff we are
halfway to success.”
“Will Abner Fownes back the man you pick? Go to the Governor fer
ye?”
She looked at him briefly, moved a step closer, and lowered her
voice.
“Abner Fownes,” she said, “is the man I believe to be the chief of the
rum runners. I believe he gave the word to kill Sheriff Churchill.”
Whitefield moved for the first time. He ran a hand through his beard
and drew a breath like a sigh.
“Life insured?” he asked.
“You will be my life insurance.”
She took him by surprise; his features actually changed for an
instant. “Me?” he said.
“When you are sheriff,” she said.
“I’m fifty. By mindin’ my business I got twenty-odd year to live.”
“He was your friend,” she said.
There was a long silence while she watched his face, and he,
looking over the top of her head, stared at the field and woodland
stretching to the horizon.
“He was my friend,” said Jared Whitefield.
“Then you will?”
“Can’t be done. Fownes has the say.”
“I think it can be done. Will you let me try?”
He considered in his ponderous way. Then he turned without a word
and walked away. He proceeded half a dozen steps and then halted.
“Yes,” he said, over his shoulder, and continued on his way.
CHAPTER XVII
CARMEL walked back rapidly, but her pace did not interfere with the
activities of her mind. She had many things to reflect upon, and not
the least of these was a sudden realization that Evan Bartholomew
Pell had, of a sudden, as it were, taken command. It was he, rather
than herself, who had risen to the emergency. He had seen the
necessities of the situation. He had comprehended the situation itself
as she had never done. While she had been obeying impulse he had
been acting intelligently. It was true he seemed to have little tangible
evidence to work upon, but, somehow, she felt he would be able to
find it. The amazing thing was that, without effort, without seeming to
do so, he had moved her into secondary place. He had told her what
to do, and she had done it without question.... Evan was a surprising
person, a person of submerged potentialities. She wondered just
what kind of man he would be if he ever came to himself and came
into his own personality. In addition to which, Carmel, like all other
women, could not but give careful consideration to a man who had
declared his love for her.
Then there was Jared Whitefield to appraise. She liked him, but
found herself somewhat in awe of his granite impassivity. She felt he
had looked through and through her, while she had not been able to
penetrate the surface of him. She had talked; he had listened. He
had made his decision, and wholly without reference to herself, or to
what she had said to him. But, on the other hand, he seemed to
have washed his hands of the responsibility for his appointment as
sheriff. If it could be managed—well and good. He would serve. But
that seemed to be all. He offered no assistance, no suggestion. He
had said “Yes” and walked out of the boundaries of the matter.
Jared Whitefield was a personality, of that she was certain. He was a
man to impress men, a man to rule, a man never to be overlooked....
Why, she wondered, had he remained inactive in Gibeon. Apparently
he had rested like a block of granite beside a busy thoroughfare,
negligent of the bustle of passing traffic. What, she wondered, did
Gibeon think of Jared. How would he appeal to Gibeon as its
candidate for sheriff?
She reached the office and found Evan Pell waiting for her.
“Well?” he said.
“I’ve found the man, and he has agreed to serve.”
“What man?”
“Jared Whitefield.”
He nodded, almost as if he had known it from the beginning. It
irritated her.
“You’re not surprised at all,” she said sharply.
“No.”
“Why?”
“Because it would have required colossal stupidity to choose any
other man—and you are not stupid.”
She looked at Evan with curiosity, and he sustained her gaze. He
was changed. She saw that he had been changing through the days
and weeks, gradually, but now he seemed to have made some great
stride and reached a destination. He did not look the same. His face
was no longer the face of an egoistic pedant. It was not alone the
laying aside of his great, round spectacles. The thing lay rather in his
expression and in his bearing. He seemed more human. He seemed
larger.... She was embarrassed.
“The petition,” she said. “I must have that.”
“Signatures would be easy to get. There are a hundred men who
would sign any petition with Jared Whitefield’s name on it. Men of
standing. But to approach one man who would go to Abner Fownes
with the story—well——” he shrugged his shoulders. “I don’t
suppose one man in a hundred realizes what is going on under the
surface in Gibeon.”
“We must take the risk.”
“I’ll prepare the petitions and have Tubal print them—at once.”
She sat down at her desk and wrote a moment, then got up and
walked with steady steps into the composing room. Evan Pell stood
looking after her with a queer expression; it was a look of loneliness,
of yearning, of self-distrust, of humility. He was thinking about Evan
Pell and of what a failure he had made in the handling of his life. He
was considering how little he knew, he who had fancied himself of
the wisest. He weighed the value of book knowledge against the
value of heart knowledge, and found himself poverty-stricken.... It
seemed so hopeless now to turn himself into the sort of man he
wanted to be; the sort of man he had come to comprehend it was
worth his while to be.
“I never would have found it out,” he said to himself, “if I had not
loved her.”
The door opened stealthily and a barefoot urchin entered whose
clothing consisted of trousers many sizes too large and a shirt so
dirty and torn as not to resemble a garment at all. He glared at Evan
and snarled:
“Where’s she?”
“Where’s who?” said Pell.
“The editin’ woman.”
“What do you want of her?”
“None of your business.... Hey, leggo of me, damn you! I’ll bite ye!
Leggo!”
Pell had the child by the nape of the neck and held him so he could
not escape. He noticed a paper crumpled in one grimy hand and
forced the fingers open. It fell to the floor, and as he reached for it
the boy wriggled free and darted out to the sidewalk, where he
grimaced horribly and twiddled his fingers at his nose. “Ya-aaa-ah!”
he squealed, and fled down the street.
Pell smoothed out the paper and read, in cramped, printed letters.
They hain’t treated me square and I’m getting even.
They’re fetching it in to-night. Truckloads. You can git
evidence at the Lakeside. Eleven o’clock.
That was all, no signature, nothing to indicate the identity of the
writer. Evan folded the paper and thrust it into his vest pocket. He
looked through the door of the composing room and frowned. The
line of his mouth was straight and narrow. Eleven o’clock, at the
Lakeside Hotel!... Queerly enough, the thought flashed into his mind.
What drew Sheriff Churchill out of his house on the night of his
disappearance?... Evan passed through the swinging gate and sat
down at his table just as Carmel re-entered the room.
“Who was in?” she asked.
“Nobody,” said Evan Pell. “Just a kid asking for blotters.”
She would go to the Lakeside Hotel. It was not in her character to do
otherwise. She would go, she would place herself in peril. Had the
note come into her hands, he had no doubt she would have
concealed it and have gone alone.... Well, she did not receive it. She
would not go. That much was sure.
Carmel spoke. “There goes Abner Fownes,” she said, and, turning,
he saw the well-known equipage with the coachman on the front
seat and Fownes, pompous, making a public spectacle for the
benefit of an admiring public, bolt upright in the rear seat.
“He’s going some place,” said Carmel. “See. He has a bag.”
“Yes,” said Pell. He remembered that Fownes had been absent from
Gibeon on the night Churchill had disappeared. “Yes, he’s going
some place.”
They watched the equipage until it disappeared, making the turn
toward the railroad station.
“Tubal will have the petitions in ten minutes,” she said. “How will we
go about getting signatures?”
“I don’t think that matters,” he said, absently.
“What?”
“I—I beg your pardon.... Er—signatures. Of course. Signatures.”
“What ails you, Mr. Pell. Of course, signatures. We weren’t speaking
of potatoes.”
His manner was strange, she thought. He seemed a trifle pale. Was
he ill?... No, he said, he was not ill, he was afraid he had been a trifle
absent-minded. Carmel eyed him sharply. The thing did not look like
absent-mindedness to her.
He arose and went to the telephone. “Give me the station, please,”
he said, and then waited. “Is this the station? This is the Free
Press.... Yes.... No news? Um!... Just saw Mr. Fownes going past
with a bag. Thought he might be going away. We like to print
something when people go away.... Bought his ticket?... To the
capital, eh?... Thank you.” He hung up the receiver, and there was a
look of profound relief on his face. This was surprising to Carmel.
Why he should be relieved by learning Fownes was on his way to
the capital was beyond her comprehension.
“Miss Lee,” he said, “there will be no time to get signers to a
petition.”
“Why?”
“Because you must start at once for the capital.”
“But the train is leaving. It will be gone before I can get to the depot.”
“Abner Fownes is going to see the Governor,” he said. “There can be
but one reason for it. He has decided he needs a sheriff. He’s
gone.... It is a six-hour trip by rail, with the change at Litchfield.”
“What of it?”
“By automobile one can make it in five hours—or less.”
“But——”
“If you will go to your hotel, please, and dress and pack a bag, I will
have a car waiting for you here.”
She frowned. This was giving orders with a vengeance.
“I’m still owner of this paper,” she said.
“Please, Miss Lee,” he said, and there was humility, pleading in his
voice. “Don’t be unreasonable now. This must be done. Nobody can
do it but you. Please, please make haste.”
She did not want to obey. It was her desire to rebel, to put him once
for all in his old subordinate place, but she found herself on her feet
in obedience. He compelled her. He had power to force her
obedience. She was amazed, angered.
“I shan’t——” she began, in a final effort to mutiny.
“Miss Lee,” he said, gravely, gently, and she was touched and
perplexed by the gentleness of his voice, “you have spoken to me of
service, of forgetting oneself to be of service to others.... Please
forget yourself now. You are not doing this for me or for yourself.... It
is necessary.... I beg of you to make haste.”
There could be no refusal. She passed through the gate and found
herself walking with rapid, almost unladylike strides, to the hotel. Up
the stairs she rushed and into her room. In five minutes she was
redressed in a gray tailored suit. Then she set about packing her
bag, and, singularly enough, the first thing she put into it was an
evening gown, the gown which she had worn but once, and that to
the final ball at the time of her graduation. Why she included this
dress she could not have said, unless feminine vanity were at work
—a hope that an opportunity to wear it might present itself.
In fifteen minutes she re-entered the Free Press office. A touring car
stood at the door, with a young man, strange to her, behind the
wheel.
“I’m ready,” she said to Evan Pell.
“Thank you,” he said, quietly. Then: “Don’t let anything prevent you
from coming to the Governor. You will know what to say. See him
before Abner Fownes gets his ear ... and ... and come back safely.”
His voice dropped, became very low and yearning, as he spoke
these final words. “Come back safely—and—try not to think of me as
—harshly as you have done.”
“I—have never thought of you harshly,” she said, affected by his
manner.
He smiled. “I am very glad I have loved you,” he said. “Will you
please remember I said that, and that it came from my heart.... It is
the one fine thing which has come into my life.... It might have
changed me—made me more as you would—less the man you have
criticized.”
“Why, Mr. Pell!... You speak as if I were never to see you again. I
shan’t be gone more than a day.”
He smiled, and there came a day not far distant when she
remembered that smile, when it haunted her, accused her—and
gave her a strange happiness.
“One never knows,” he said, and held out his hand. She placed her
hand in his, and then he performed an act so out of tune with Evan
Pell, pedant and egoist, that Carmel gasped. He lifted her hand to
his lips. The gesture was not artificial, not funny. There was a grave
dignity, a sincerity in the act which made it seem quite the right thing
to have done. “Good-by,” he said. “You are very lovely.... Please
make haste....”
He helped her into the car, and she turned. “Mr. Pell——” she said,
but he was gone, had returned to the office and was invisible.
“Ready, miss?” the driver asked.
“You know where you are to go?”
“Yes, miss.”
“Whose car is this?”
“Mr. Whitefield’s,” said the driver, as he threw in his gear and the
machine moved up the street.
Carmel’s mind was not on the car, nor on its destination, nor upon
her errand. It was upon Bartholomew Pell.... Could she have seen
him seated before his table, could she have read his thoughts, have
comprehended the expression of happiness upon his face, she
would have thought even more urgently of him.... For he was saying
to himself: “Thank God she’s out of it. She’s safe. I’ve done that
much, anyhow.”
He drew the mysterious note from his pocket and studied it
attentively. “She would have gone,” he said, “so I shall go....
Doubtless it is a trap of some sort—but it may not be.... And she is
safe—she is safe.”
CHAPTER XVIII
CARMEL wondered how one went about it to obtain a private
interview with a Governor. She was still young enough and
inexperienced enough in life’s valuations to regard a man in that
position as necessarily above the ordinary run of men. His office
invested him with a certain glamour, a fictitious greatness.
Governors, Senators, Presidents! Youth invested them with a terrific
dignity. It is somewhat difficult, even for the wise and prudent, to see
the man apart from his vestments; to understand that there is, in
reality, very slight difference between human beings, and to
approach those in authority with the sure knowledge that, no matter
how lofty their position, they have, at best, but two arms and two
legs, a fondness for mince pies, and a failing for colds in the nose.
Governors quarrel with their wives, and have ingrowing toe nails.
The forty-eight of them, heads of the several states of the Union,
remind one of the main street in a boom town—two stories on the
sidewalk, but a ramshackle shed in the rear....
No sooner did the dome of the Capitol appear through a break in the
wheels than Carmel began to dress herself mentally for the meeting.
She had a horrible fear she would become tongue-tied and thrust her
thumb in her mouth like an embarrassed little girl who has forgotten
her piece.... She glanced at her watch. It was five o’clock.
How late did Governors work at governing?... She directed her
chauffeur to drive to the Capitol, and there she alighted because she
had no idea what else to do. She climbed the imposing steps and
entered the building. It was a repellent sort of place; a mausoleum of
assassinated ambitions, and it chilled her. The corridors were all but
deserted.
Leaning against a column adjacent to a brass cuspidor was an old
man in a uniform which might have been that of a prison guard, a
janitor, or a retired street car conductor. Carmel approached him.

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Nanomaterials for food applications

MARTA MARTINEZ SANZ

For information on all Elsevier publications visit our website at

Publisher: Matthew Deans

1. Nanomaterials for Food Applications: General Introduction and Overview

3. Nanoencapsulation: Techniques and Developments for Food Applications 35

4. Nanostructured Minerals and Vitamins for Food Fortiﬁcation and Food

Part 2: Nanotechnologies for Food Processing

Part 3: Nanosensors for Food Quality and Safety

7. Nanoparticles as Biosensors for Food Quality and Safety Assessment 147

Part 4: Nanotechnologies for Food Packaging

8.1 Introduction 205

9. Bioactive Packaging: Combining Nanotechnologies With Packaging

9.5 Bioactive Packaging Containing Nanostructures 256

Part 5: Nanotoxicology and Regulatory Status

10.1 Introduction 273

11. Characterization of Nanomaterials: Tools and Challenges 313

12. Toxicological Hazard Analysis of Nanomaterials With Potential for

13. Regulatory Status of Nanotechnologies in Food in the EU 381

combining these active/bioactive substances included in the packaging structure with

Nanomaterials for Food Applications:

Nanomaterials for Food Applications © 2019 Elsevier Inc.

1.2 POTENTIAL OF NANOTECHNOLOGIES TO IMPROVE CONSUMER

1.3 NANOTECHNOLOGIES FOR FOOD PROCESSING AND SENSING

1.4 FOOD PACKAGING NANOTECHNOLOGIES

1.5 CHARACTERIZATION, TOXICOLOGICAL ASSESSMENT, AND

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MARTA MARTINEZ SANZ