Advances in the Determination of Xenobiotics in Foods -  - E-Book

Advances in the Determination of Xenobiotics in Foods E-Book

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Determining the presence of different types of toxic compounds (or xenobiotics) in food requires precise analytical methodologies. Examples of these techniques include separation techniques coupled to mass spectrometry, Variations in methods used depend on the physicochemical properties of each xenobiotic being tested for.
Advances in the Determination of Xenobiotics in Foods explains recent developments in the field of xenobiotic determination in food. Readers are introduced to xenobiotic testing techniques through extensive reviews. Chapters also cover details about contaminants coming from food contact materials (such as plasticizers, food additives, polymer monomers/oligomers and non-intentionally added substances), substances used for food processing and sensing (nanoparticles), and residues of pesticides (that can also be present in the final food product). The book also includes information about specific xenobiotics that, due to their global distribution in the environment, are also likely to enter the food chain. Some of them are regulated (persistent organic pollutants and heavy metals) but there are many other types of contaminants (halogenated flame-retardants, perfluorinated compounds and micro- and nanoplastics) that must also be controlled. In addition, some xenobiotics could be present in the final food consumed because of food treatments (acrylamide, furan, heterocyclic aromatic amines, and glycidol esters). Finally, the concluding chapters of the book are devoted to the presence of natural contaminants such as mycotoxins and biogenic amines.
The combination of extensive information of analytical techniques for xenobiotics along with a categorical treatment of food contaminants makes this volume a handy reference for food science and technology students and technicians involved in food safety and processing management roles.

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Table of Contents
Welcome
Table of Content
Title
BENTHAM SCIENCE PUBLISHERS LTD.
End User License Agreement (for non-institutional, personal use)
Usage Rules:
Disclaimer:
Limitation of Liability:
General:
PREFACE
DEDICATION
List of Contributors
Safety Assessment of Active Food Packaging: Role of Known and Unknown Substances
Abstract
INTRODUCTION
Antioxidant Packaging
Types of Antioxidant Packaging
Oxygen Scavengers
Free Radical Scavengers
Antimicrobial Packaging
Antimicrobials
Safety Assessment
Active Substances
Food Contact Materials
CONCLUDING REMARKS AND FUTURE TRENDS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Microplastics and Nanoplastics in Food
Abstract
INTRODUCTION
MICROPLASTICS AND NANOPLASTICS DEFINITION
MICROPLASTICS AND NANOPLASTICS COMPOSITION
Polymers
Flame-Retardants
Plasticizers
Antioxidants and Stabilizers
PRESENCE OF MICROPLASTICS AND NANOPLASTICS IN FOOD
Fisheries and Aquaculture Products
Salt
Water
METHODS OF CHARACTERIZATION AND DETECTION OF MICROPLASTICS
Sampling
Pretreatment and Purification
Separation
Separation by Density
Filtration
Digestion
Dissection and Cleaning
Homogenization
Digestion
Identification
Visual Identification
Scanning Electron Microscope (SEM)
Detection and Quantification
FTIR and ATR
Raman Spectroscopy
Pyr-GC-MS
Quantification of Microplastics
METHODS OF ANALYSIS FOR CHEMICAL COMPOUNDS RELATED TO MICROPLASTICS
Methods of Analysis for Plastics Additives
Brominated Flame Retardants (BFRs)
Phthalates
Bisphenol A and Nonylphenols
Methods of Analysis for Plastics Contaminants (Dioxins, PCBs…)
MICROPLASTICS DIETARY INTAKE
LIMITATIONS FOR FOOD SAFETY RISK ASSESSMENT
Examples of Risk Assessment
INVESTIGATION GAPS
Analytical Control and Quality Assurance
Sample Contamination
Sampling
Visual Identification of Microplastics
Detection of micro-Plastics by Different Analytical Techniques
CONCLUDING REMARKS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENT
REFERENCES
Nanotechnology in the Food Field: Application of Metal-Based Nanoparticles
Abstract
INTRODUCTION
METAL-BASED NANOPARTICLES IN THE FOOD SECTOR
Nanoparticles in Food Processing
Nanoparticles as Antimicrobial Agents: Food Safety
Nanoparticles in Food Packaging
Nanoparticles for Food Sensing
Nano-food Regulatory Issues in the European Union
CHARACTERIZATION OF METAL-BASED NANOPARTICLES: THE NEED FOR USING A MULTI-TECHNIQUE APPROACH
Factors Affecting Stability of Nanoparticles
Nanoparticles Migration from Packaging to Food
CONCLUDING REMARKS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Halogenated and Organophosphorus Flame Retardants
Abstract
INTRODUCTION
FLAME RETARDANTS
Polybrominated Diphenyl Ethers and Hexabromocyclododecane
Emerging Flame Retardants
Organophosphorus Flame Retardants
Physicochemical Properties
Bioaccummulation and Biomagnification
Toxicity
Legislation
ANALYTICAL METHODOLOGIES
Main Compounds
Extraction
Lipid Removal
Clean-up
Instrumental Analysis
Quality Parameters
LEVELS IN FOOD AND INTAKE
CONCLUDING REMARKS
LIST OF ABBREVIATIONS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Dioxins and PCBs in Food and Feed Matrices: Advances in Physico-Chemical Methods and EU Regulatory Framework
Abstract
INTRODUCTION
ADVANCES IN EXTRACTION AND PURIFICATION TECHNIQUES
ADVANCES IN MASS SPECTROMETRY TECHNIQUES
GC-HRMS as the Reference Technique
Time-of-Flight Mass Spectrometry (ToF-MS)
Tandem Mass Spectrometry with Ion Trap Analyzer (IT-MS/MS)
Tandem Mass Spectrometry with Triple Quadrupole Configuration (QqQ-MS/MS)
New HRMS Techniques
CONCLUDING REMARKS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENT
REFERENCES
Pesticides
Abstract
INTRODUCTION
LEGISLATION
ANALYTICAL METHODS
Extraction
Solvent Extraction (SE)
Matrix Solid-Phase Dispersion (MSPD)
Head-Space Solid Phase Micro-Extraction (HS-SPME)
Clean-up
Liquid-Liquid Partitioning (LLP)
Solid-Phase Extraction (SPE)
Dispersive Solid-Phase Extraction (dSPE)
Gel Permeation Chromatography (GPC)
Determination Techniques
Chromatographic Methods
Gas Chromatography (GC)
Liquid Chromatography (LC)
Other Chromatographic Techniques
Ambient Mass Spectrometry (AMS)
Other Spectroscopic Techniques
Immunoassays
Sensors
CONCLUDING REMARKS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENT
REFERENCES
Perfluoroalkyl Substances (PFASs) in Foodstuffs and Human Dietary Exposure
Abstract
INTRODUCTION
Sources of Human Exposure to PFASs
Select Exposure Related Epidemiological Studies of PFASs
METHODS
Food Sample Collection
PFAS Analysis in Food
PFAS Extraction in House Dust
PFAS Analysis in Water
Method Validation
Instrumental Analysis
Background Contamination
Matrix Effects and Internal Standards
Instrumental Performance and Limit of Quantification (LOQ)
HUMAN EXPOSURE ASSESSMENT OF PFASs
External/Environmental Sources Method
Internal Dose/Biomonitoring Method
RESULTS
PFAS Concentrations in Food and Beverage Samples
PFASs in Tap Water
PFASs in Indoor Dust
Human Exposure to PFASs through Environmental Exposures
Assessment of Exposure to PFOS
PFOS Exposure in Infants below 1-Year Old
PFOS Exposure in Children, Adolescents, and Adults
PFOS Exposure Dose Calculation from Biomonitoring Data
Assessment of Exposure to PFOA
PFOA Exposure to Infants Below 1 Year Old
PFOA Exposure in Children, Adolescents, and Adults
PFOA exposure dose calculation from biomonitoring data
CONCLUDING REMARKS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Mercury
Abstract
INTRODUCTION
FOOD MATRICES WHERE Hg IS OFTEN DETERMINED
ADVANCES ON THE DETERMINATION OF TOTAL Hg IN FOOD
Sample Treatment for Total Hg Determination Assisted by Solid-Phase Extraction
RECENT ADVANCES IN SPECIATION ANALYSIS OF MERCURY IN FOOD
Hg Speciation in Food by Using High Pressure Liquid Chromatography
Chromatographic Separation Assisted by Solid-Phase Extraction
Trends in the Choice of Stationary and Mobile Phases in HPLC for Hg Analysis
Vapor Generation of Hg Species Assisted by Nanomaterials
Hg Speciation in Food by Non-Chromatographic Methods
Non-Chromatographic Speciation Assisted by SPE
Non-Chromatographic Methods Assisted by Nanomaterials
ADVANCES AND TRENDS IN THE USE OF Hg STABLE ISOTOPES
CONCLUDING REMARKS
LIST OF ABBREVIATIONS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Process Contaminants
Abstract
INTRODUCTION
General Considerations to Food Safety
Risk Assessment Scheme
Estimation of Food Chemical Intake
Toxicological Considerations
Process Contaminants
ACRYLAMIDE
Characterization and Toxicology
Chemical Formation in Foods
Occurrence and Exposure
Factors Affecting the Acrylamide Formation
Acrylamide Precursors
Time and Temperature of the Process
Water Activity and pH
Other Factors
Exposure
Mitigation
Levels of Precursors
Process Parameters
Other Factors Removing or Trapping Acrylamide
Methods of Analysis
FURAN
Characterization and Toxicology
Chemical Formation in Foods
Occurrence and Exposure
Mitigation
Preventive Strategies
Removal Strategies
Methods of Analysis
HETEROCYCLIC AROMATIC AMINES (HAAs)
Characterization and Toxicology
Chemical Formation in Foods
Occurrence and Exposure
Mitigation
Methods of Analysis
CHLOROPROPANOLS AND ESTERS, GLYCIDOL AND GLYCIDYL FATTY ACID ESTERS
Characterization and Toxicology
3-MCPD and its Fatty Acid Esters
Glycidol and Glycidyl Esters
Chemical Formation in Foods
Occurrence and Exposure
Mitigation
Legislation
Methods of Analysis
Direct Determination
Indirect Determination
CONCLUDING REMARKS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENT
REFERENCES
Mycotoxins
Abstract
GENERAL INTRODUCTION
Preamble
OVERVIEW
MAIN MYCOTOXINS
Mycotoxins Produced by Aspergillus and/or Penicillium Fungi
Aflatoxins
Production
Presence in Food
Toxicology
Ochratoxin A
Production
Presence in Food
Toxicology
Patulin
Production
Presence in Food
Toxicology
Mycotoxins Produced by Fusarium Fungi
Fumonisins
Production
Presence in Food
Toxicology
Trichothecenes
Production
Presence in Food
Toxicology
Zearalenone
Production
Presence in Food
Toxicology
Emerging Fusarium Mycotoxins
Production
Presence in Food
Toxicology
Mycotoxins Produced by Alternaria Fungi
Production
Presence in Food
Toxicology
LEGISLATION
MYCOTOXIN ANALYSIS
Sampling
Extraction and Purification
Confirmation Techniques
Chromatographic Techniques
Gas Chromatography
Liquid Chromatography
Mass Spectrometry
EXPOSURE ASSESSMENT
METABOLISM AND BIOMARKERS
FUTURE TRENDS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Biogenic Amines
Abstract
INTRODUCTION
Meat
Seafood
Cheese
Beer and Liqueurs
Wine
Baby Foods
EXTRACTION AND PURIFICATION OF BAs IN FOOD
ANALYTICAL TECHNIQUES IN BAs ANALYSIS
Liquid Chromatography (LC)
Gas Chromatography (GC)
Capillary Electrophoresis (CE)
Others Methodologies
CONCLUDING REMARKS
CONSENT FOR PUBLICATION
CONFLICT OF INTEREST
ACKNOWLEDGEMENTS
REFERENCES
Current and Future Developments in Food Science
(Volume 1)
Advances in the Determination of Xenobiotics in Foods
Edited by
Belén Gómara
Department of Instrumental Analysis and Environmental Chemistry,
Institute of General Organic Chemistry (IQOG),
Spanish National Research Council (CSIC),
Madrid,
Spain
María Luisa Marina
Department of Analytical Chemistry, Physical Chemistry and Chemical Engineering,
University of Alcalá Alcalá de Henares (Madrid),

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PREFACE

Xenobiotics had been and presently are of great concern, both for the society and the health authorities all over the world. Xenobiotics in food may include a huge variety of compounds of different nature. Nowadays, one of the groups that have caught the attention of researchers and authorities are food chain residues. These compounds are chemicals unintentionally present in the food due to the different procedures of production and preparation methods to which foodstuffs are subjected. Among them, compounds related to food contact materials such as plasticizers and plastic monomers are one of the xenobiotics mostly supervised by the European Food Safety Authority (EFSA) and the Environmental Protection Agency (EPA) and Food and Drug Administration (FDA) in the United States. In addition, pesticides can also be present in the final foodstuff because of their previous use in the field or on the farm. On the other hand, due to the global distribution of environmental pollutants, they are also susceptible to end up in the food chain because of different processes of deposition and/or bioaccumulation. There are several classes of environmental pollutants. Some of them are regulated by local or global legislations such as persistent organic pollutants and heavy metals. There are also many other emerging contaminants that must be controlled such as some halogenated flame retardants and perfluorinated compounds, among others. In addition, some xenobiotics could be present in the final food consumed as a result of food treatments, as is the case of acrylamide and furan which are related to high-temperature cooking processes. Finally, the presence of natural contaminants such as mycotoxins, aflatoxins and biogenic amines in the final foodstuffs must be controlled too.

The control of all these compounds would not be possible without the development of advanced analytical methodologies enabling their unequivocal, precise and accurate determination in foodstuffs. In this regards, one of the most employed methodologies is the separation techniques coupled to mass spectrometry. Depending on the physicochemical properties of each xenobiotic, gas (GC) or liquid (LC) chromatography can be applied for its separation, identification and quantification. Constant research is being carried out in order to develop more sensitive and selective methods for the determination of these xenobiotics at the low concentration levels they use to be present in foodstuffs. Novel analytical approaches in this field are fast GC and ultra-high performance LC (UHPLC) which have been successfully applied to study some of these xenobiotics. In addition, highly sensitive and selective mass analyzers such as triple quadrupole, Orbitrap or other hybrid systems combining some of them and novel developments such as ion mobility equipment are being recently applied to these purposes. These advances in combination with fast and environmentally friendly sample extraction and purification methods provide the society and authorities with the necessary methods for controlling and regulating, if necessary, the presence of all these xenobiotics in food.

Therefore, this book is aimed to present some of the most recent advances and developments achieved in the determination of different xenobiotics in foods. Chapters are organized according to the type of xenobiotic under study.

This book was inspired by the context of the AVANSECAL-CM and AVANSECAL-II-CM research projects funded by the Comunidad of Madrid and European FEDER program and headed by Professor María Luisa Marina from 2014 to nowadays, which was the continuation of two previous research projects (ANALISYC and ANALISYC-II) headed by Professor María José González from 2006 to 2013. Along all these years, a numerous group of researchers made considerable efforts to develop innovative analytical methodologies to control and improve food quality and safety with very relevant results in this field which have been and are being recognized at the international level. The editors are very grateful to these researchers, especially to those who have contributed to this e-book, and dedicated this e-book to Professor María José González in her retirement as a worm acknowledgement for her valuable contribution in the field of xenobiotics analysis.

Experts and researchers in analytical chemistry, food safety and xenobiotic analysis and newcomers in these fields such as Ph.D. students or chemists working in control laboratories or laboratory technicians will find in this e-book updated information including a set of advanced analytical methods used for the analysis of a broad spectrum of xenobiotics revealing the most interesting features and drawbacks to be overcome in this field. PhD students will learn more about novel analytical developments, they will acquire knowledge about xenobiotics and know in depth the field of food contamination. Finally, chemists working in control laboratories or laboratory technicians will have a very useful tool to face the problems arising on food safety.

We are very grateful to all the authors for their relevant contributions to this e-book.

Efforts in this field will be pursued in the next four years, thanks to the fundings from the Comunidad of Madrid (Spain) and European FEDER program through the new AVANSECAL-II-CM research project.

Belén Gómara Department of Instrumental Analysis and Environmental Chemistry Institute of General Organic Chemistry (IQOG) Spanish National Research Council (CSIC) Madrid SpainMaría Luisa Marina Department of Analytical Chemistry, Physical Chemistry and Chemical Engineering University of Alcalá Alcalá de Henares (Madrid) Madrid Spain

DEDICATION

Dedicated to Professor Maria José González Carlos in her retirement

List of Contributors

Alberto RitieniUniversità di Napoli Federico II, Department of Pharmacy, Via D. Montesano, 49 -, 80131 Napoli, ItalyBeatriz Gómez-GómezDepartamento de Química Analítica. Facultad de Ciencias Químicas. Universidad Complutense de Madrid. 28040. Madrid, SpainCristina NerínI3A – Aragón Institute of Engineering Research, University of Zaragoza, Calle María de Luna 3, 50018 Zaragoza, SpainEsther Garrido-GamarroFood Safety and Quality Officer, Fisheries and Aquaculture, Department, Food and Agriculture Organization of the United Nations (FAO). Viale delle Terme di Caracalla, 00153 Roma RM, ItalyEsteban AbadLaboratorio de Dioxinas. Departamento de Química Ambiental. IDAEA-CSIC. C/ Jordi Girona, 18-26, 08034 Barcelona, SpainEthel EljarratInstitute of Environmental Assessment and Water Research, Spanish National Research Council (IDAEA-CSIC). C/ Jordi Girona, 18-26, 08034 Barcelona, SpainFilomena SilvaARAID – Agencia Aragonesa para la Investigación y el Desarollo, Av. de Ranillas 1-D, planta 2ª, oficina B, 50018 Zaragoza, Spain Faculty of Veterinary Medicine, University of Zaragoza, Calle de Miguel Servet 177, 50013 Zaragoza, SpainFrancisca HolgadoInstitute of Food Science, Technology and Nutrition, Spanish National Research Council (ICTAN-CSIC), C/ José Antonio Novais, 10, 28040 Madrid, SpainFrancisco J. MoralesInstitute of Food Science, Technology and Nutrition, Spanish National Research Council (ICTAN-CSIC), C/ José Antonio Novais, 10, 28040 Madrid, SpainGianni SagratiniUniversity of Camerino, School of Pharmacy, Via S. Agostino 1, 62032 Camerino (MC), ItalyGiovanni CaprioliUniversity of Camerino, School of Pharmacy, Via S. Agostino 1, 62032 Camerino (MC), ItalyJordi PareraLaboratorio de Dioxinas. Departamento de Química Ambiental. IDAEA-CSIC. C/ Jordi Girona, 18-26, 08034 Barcelona, SpainKurunthachalam KannanWadsworth Center, New York State Department of Health, and Department of Environmental Health Sciences, University at Albany, State University of New York, Albany, NY 12201, USAManuela ÁbalosLaboratorio de Dioxinas. Departamento de Química Ambiental. IDAEA-CSIC. C/ Jordi Girona, 18-26, 08034 Barcelona, SpainMarta MesíasInstitute of Food Science, Technology and Nutrition, Spanish National Research Council (ICTAN-CSIC), C/ José Antonio Novais, 10, 28040 Madrid, SpainMassimo RicciutelliUniversity of Camerino, School of Pharmacy, Via S. Agostino 1, 62032 Camerino (MC), ItalyÒscar Aznar-AlemanyInstitute of Environmental Assessment and Water Research, Spanish National Research Council (IDAEA-CSIC). C/ Jordi Girona, 18-26, 08034 Barcelona, SpainPilar Fernández-HernandoDepartamento de Ciencias Analíticas, Facultad de Ciencias, Universidad Nacional de Educación a Distancia (UNED). Paseo Senda del Rey nº 9, 28040, Madrid, SpainQian WuWadsworth Center, New York State Department of Health, and Department of Environmental Health Sciences, University at Albany, State University of New York, Albany, NY 12201, USARaquel BecerrilI3A – Aragón Institute of Engineering Research, University of Zaragoza, Calle María de Luna 3, 50018 Zaragoza, SpainRosa Mª Garcinuño-MartínezDepartamento de Ciencias Analíticas, Facultad de Ciencias, Universidad Nacional de Educación a Distancia (UNED). Paseo Senda del Rey nº 9, 28040, Madrid, SpainSauro VittoriUniversity of Camerino, School of Pharmacy, Via S. Agostino 1, 62032 Camerino (MC), ItalyVicente AndreuEnvironmental and Food Safety Research Group of the University of Valencia (SAMA-UV), Desertification Research Center (CIDE), Joint Research Center CSIC-UV-GV, Moncada, SpainYolanda PicóEnvironmental and Food Safety Research Group of the University of Valencia (SAMA-UV), Desertification Research Center (CIDE), Joint Research Center CSIC-UV-GV, Moncada, SpainYelko Rodríguez-CarrascoUniversity of Valencia, Department of Food Chemistry and Toxicology, Av/ Vicent A. Estellés, s/n 46100 Burjassot,, Valencia, SpainYolanda MadridDepartamento de Química Analítica. Facultad de Ciencias Químicas. Universidad Complutense de Madrid. 28040. Madrid, SpainZoyne Pedrero ZayasCNRS/UNIV PAU & PAYS ADOUR, Institut des Sciences Analytiques et de Physicochimie pour l’Environnement et les Matériaux, UMR 5254, 64000, Pau, France

Safety Assessment of Active Food Packaging: Role of Known and Unknown Substances

Filomena Silva1,2,Raquel Becerril3,Cristina Nerín3,*
1 ARAID – Agencia Aragonesa para la Investigación y el Desarollo, Av. de Ranillas 1-D, planta 2ª, oficina B, 50018 Zaragoza, Spain
2 Faculty of Veterinary Medicine, University of Zaragoza, Calle de Miguel Servet 177, 50013 Zaragoza, Spain
3 I3A – Aragón Institute of Engineering Research, University of Zaragoza, Calle María de Luna 3, 50018 Zaragoza, Spain

Abstract

Nowadays, consumers are more aware of what they eat and also request, minimally processed foods and they tend to prefer biodegradable or bio-based packaging. One of the most accepted technologies to battle this problematic is active packaging. Active packaging protects the food product by extending its shelf-life while guaranteeing its safety through the addition of antimicrobials or antioxidants that actively interact with the packaging atmosphere or the food product to avoid oxidation processes, microbial growth and other routes responsible for food spoilage. Although yet not fully implemented in Europe, active packaging is expected to reach a compound annual growth rate of 6.9% in 2020. However, in order to get these active packaging solutions into the market, their safety must be ensured and they must comply with the European legislation on the topic, both for the active substances incorporated into the packaging materials as for the packaging material itself. These packaging materials, either plastic or bio-based, can pose food safety risks to consumers due to the migration of compounds from the packaging to the food product. Compounds like plasticizers, additives, polymer monomers/oligomers and even non-intentionally added substances (NIAS) can migrate from the packaging material to the food product at concentrations capable to endanger human health and, therefore, they must be correctly detected and identified, to allow a correct risk assessment and strict monitoring of the packaging materials available.

Keywords: Active packaging, Antioxidant, Antimicrobial, Migration, Release, Food contact materials, Bio-based polymers, Natural compounds, Non-intentionally added substances.
*Corresponding author Cristina Nerín: Aragón Institute of Engineering Research, Campus Rio Ebro, Edificio Torres Quevedo, Universidad de Zaragoza, Calle Maria de Luna 3, 50018 Zaragoza, Spain; Tel: +34976761873; E-mail: [email protected]

INTRODUCTION

Nowadays, consumers are more aware of what they eat and also request minimally processed foods and they tend to prefer biodegradable or bio-based packaging over the traditional plastic ones. Therefore, there is an urgent real need to develop newer and safer food packaging systems to improve food shelf-life, whether to reduce food waste (packaged food or the package itself) [1] or to distribute products to more distant places. Furthermore, there is a growing need to provide new solutions to ensure the safety and quality of the packaged foods and products. Due to all these demands, there is a growing market for the development of new packaging solutions, called active packaging (AP), to be applied within several fields, such as pharmaceutical, healthcare or food industries [2]. Active packages are based on the incorporation of active agents with the food packages, thus avoiding the direct addition of chemicals to the food. These active agents can be either incorporated directly in the packaging materials, or be included inside a package as pads, trays, sachets or pouches. These active agents include antioxidants, antimicrobials or absorbers that hand over new properties to the pre-existent material such as oxygen or free radical scavenging (antioxidant/absorber) or microbiological control (antimicrobial) [2]. Over the last decades, active packaging has become a reality for the food packaging industry with the constant growth of the global market for active/intelligent packaging, reaching a compound annual growth rate of 6.9% [3]. There is a vast array of AP solutions currently available in the market; with the vast majority of them having the consumer as the final user and a few of them being intended for business-to-business use (Table 1).

Table 1Examples of worldwide commercially available active packaging solutions.TradenameCompanyAP TypeMaterialsCountryConsumersB2BAgeless®Mitsubishi Gas Chemical Co. Inc.Oxygen scavengerSachetsJapanXTorishigeDessicantLaminate papersJapanXEMAP/AMAPPerfotecModified Atmosphere Packaging (controlled permeability of the packaging and controlled atmosphere)Plastic traysThe NetherlandsXXFreshPaperFenugreenFiber-based sheets with organic spicesPaper sheetUSA (also export in Europe)XXMegaCO2PomonaMoisture absorbing pads combined with CO2 scavengerpadsPolandXEthenAbsorbers/ETENSachetsPomonaEthylene scavengersachetsPolandXOxyguardClariantOxygen scavengerssachetsUSAXBreatheWayLandec CorporationSelective permeability filmsfilmsUSAXDarex OSTDarexOxygen scavengercrownUSAOxyFreshSTANDA Laboratories (EMCO)CO2 scavenger / O2 emittersachetsFranceXSanocoatMondi Packaging Flexibles AGAntimicrobial paperpaperAustria/GermanyXNAErze Ambalaj/Parx PlasticsAntimicrobial trayplastic traysTurkeyXAntioxidantPackBTSAAntioxidant filmplastic filmsSpainXSupasorbThermariteHigh capacity moisture absorbing filmpadsMalaysiaXFresh-r-PaxMaxwell Chase Technologies LLCMoisture absorbenttrays, pads, pouchesUSAXDriFresh® SeaFresh™ Fresh-HoldSiraneAbsorbing pad (moisture/ice/odor) with CO2 emitterpadsUK, available in PolandXDri-Fresh® SeaFresh™ Ice-MatsSiraneSeawater-releasing pad to extend seafood shelf-lifepads, matsUK, available in PolandXDri-Fresh® Fresh-Hold™ OASiraneOdour-scavenging padpads, labelsUK, available in PolandXDri-Fresh® Fresh-Hold™ ABSiraneAntibacterial padpadsUK, available in PolandXNAArtibal S.A.Antimicrobial coatingfilmsSpainXNAArtibal S.A.Antioxidant coatingfilmsSpainXNAGoglio SpAAntioxidant filmfilmsItalyXNASAMTACKAntioxidant adhesive for multilayeradhesiveSpainXRycoat F-100, Emulactiv C-1REPSOL YPF Lubricantes & EspecialidadesAntimicrobial/antioxidant coatingpaper/cardboardSpainXXNACellCombCO2 emitterpadsSwedenX

These packages have been used to preserve all kinds of foods ranging from more perishable goods, as fresh fruit, vegetables, meat, fish and cheese to more processed foods such as breads, cakes and sweets, sauces and jams, processed and dried meat, snacks and even baby food and pet food. These AP solutions include several absorbers such as moisture and odor absorbers, and ethylene and carbon dioxide scavengers. With respect to antioxidant packaging, the two main types of AP available are the use of oxygen and free radical scavengers. When dealing with antimicrobial packaging, there is a broad array of technologies available aiming at reducing microbial growth in the food product, either by changing the atmosphere (selective permeability films for modified atmosphere and carbon dioxide emitters) or by adding antimicrobial substances ranging from chemicals to natural products such as essential oils or herb extracts. The mode of action of each AP depends on the active agent incorporated and the packaging design as well as the characteristics of the packaged food product (Table 2).

The emergence and development of these new packaging technologies that interact with food triggered a response from the authorities to ensure their safety towards the consumers. In this regard, the European Union adopted specific legislation to regulate the use and application of these new active and intelligent packaging systems, namely European Committee regulations 1935/2004 and 450/2009. Regulation (EC) No 1935/2004 on materials and articles intended to come into contact with food [4] defines for the first time active packaging and authorizes its use by establishing limits to guarantee the safety of consumers. Regulation (EC) No 450/2009 on active and intelligent materials and articles intended to come into contact with food [5] includes general requirements stated in Regulation 1935/2004/EC and provides new specific requirements for the safe use of active and intelligent packaging.

Table 2Mode of action of different active packaging technologies available on the market.Active PackagingFunctionAP MaterialMode of ActionAntioxidantOxygen scavengerIron-based sachetsThe iron contained in the sachets is prone to oxidation, absorbing oxygen in the processFree radical scavengerMulti-layer filmThis multilayer film contains green tea extract (GTE) and acts by scavenging free radicals formed in the atmosphere of the package, using catechins as free radical acceptor molecules. There is no direct interaction between antioxidant compounds and foods, as the green tea extract is crafted inside the multilayer film, with none or negligible specific migration of green tea compounds from the packageAntimicrobialCarbon dioxide emitterHighly absorbent padsThe liquids released by the packaged food activate the CO2-releasing reaction. As the majority of the bacteria present in foods are aerobic (e.g., require oxygen), the maintenance of a steady CO2 concentration causes an antimicrobial effect on the food as it inhibits aerobic bacteria growth.Antimicrobial releaserAntimicrobial multi-layer filmThese films contain natural volatile extracts The antimicrobial activity of these films is exerted in the vapor phase, taking advantage of the inherent volatility of the extracts used; therefore, no direct contact is required between the package and the food product, which allows for a more homogeneous distribution and action of the antimicrobial agent in the whole food productOtherPermeability controlPackaging polymeric filmCrystallizable polymer with temperature switch: at low temperatures, the polymer is in a more compact, crystalline structure that has low permeability; meanwhile, when temperature rises, polymer structure changes to an amorphous state which results in an increased permeabilityEthylene absorberAbsorbing padsThese pads contain substances capable of absorbing ethylene and thus prevent fruit ripeningDesiccantLaminate paperCalcium chloride is used as moisture absorber due to its hygroscopic features The absorber is included in a cellulose matrix to potentiate moisture absorption

According to these regulations, active packaging is allowed to change the composition of the atmosphere around the packaged food, providing that the active substance is an authorized compound and the originated change complies with EU regulation. Besides, AP should be correctly labelled to prevent improper use and misunderstanding by consumers. Another important point of these regulations is the establishment of standardized procedures to assess the safety of both the passive parts of active materials, such as the packaging materials, and the active compounds [6].

Antioxidant Packaging

The presence of an oxygen-rich atmosphere in the packaging headspace is one of the most critical factors affecting both food quality and shelf-life. The presence of residual oxygen can led to product oxidation resulting in changes in its organoleptic properties, such as color, odor, taste, etc [7] (Table 3); in its nutritional value due to oxidation of important vitamins, fatty acids, proteins, among others [8, 9] (Table 3); and its microbial load as oxygen promotes the growth of aerobic foodborne spoilage and pathogenic bacteria [10]. All the above mentioned detrimental effects of oxygen on food products can have a severe impact on human health, either by the consumption of microbiologically contaminated food or by the ingestion of food oxidation products. These products are formed during food production, processing and storage, being currently described as potentially harmful to humans, as they have been linked to inflammatory and to the onset of carcinogenic processes [11]. Regarding microbial contamination, aerobic pathogens such as Salmonella spp, E. coli and L. monocytogenes, highly prevalent in food products, can lead to severe foodborne illness, with significant morbidity and mortality rates [12]. Additionally, the decrease in consumer’s acceptance of oxidized foods with altered organoleptic properties and nutritional value results in great economic losses due to food waste [13]. Consequently, the food industry aims to exclude oxygen from food packaging, which is mainly performed by gas flushing or modified atmosphere packaging (MAP) processes, or more efficiently by using antioxidant packaging capable of controlling residual oxygen [14].

Table 3Sensorial and nutritional changes caused by oxygen in packaged foods.Food ProductOrganoleptic and Nutritional Changes due to Oxygen PresenceReferenceMeat productsDevelopment of rancidity Potential formation of toxic aldehydes Loss of nutritional quality because of polyunsaturated fatty acid (PUFA) degradation[15]Oils and fatsDevelopment of off-flavors and odors (such as tallowy, painty, burned, fishy, grassy) Vitamin E content of vegetable oils and the vitamin A content of fish oils are reduced, if not entirely destroyed Degradation of essential dietary fatty acid components Development of rancidity Darkening of color[16]Dairy productsLipid oxidation and formation of peroxides Development of rancid flavor Off flavors (tallowy, oily, and fishy) in butters Tallowy or oxidized flavor in ice cream Tallowness in sweetened condensed milk Formation of toxic aldehyde and ketone compounds[17]Baked productsDegradation of vitamins (A, C, D, E, and folate), essential fatty acids (linoleic acid), and essential amino acids like methionine, cysteine, histidine, lysine, and tryptophan Development of rancidity Changes in texture[18]Fruits and vegetablesPhenolic browning of fruit/vegetables Changes in texture (softening) Loss of vitamin C (ascorbic acid) Acceleration of respiration Discoloration[19]

Types of Antioxidant Packaging

Overall, we can distinguish between two main types of antioxidant compounds: primary antioxidants, such as free-radical scavengers, which target oxidation products; and secondary antioxidants, such as metal chelators, UV absorbers, singlet oxygen (1O2) quenchers and oxygen scavengers, which prevent oxidation.

Oxygen Scavengers

Oxygen may be accessible in the package through several routes such as entrance through the packaging material, minor leakage due to poor sealing and material barrier properties, on account of the residual oxygen in the air encased in the container, deficient gas flushing or oxygen evacuation from the container head-space [20]. Oxygen scavenging compounds react with the oxygen present in the package to reduce its concentration and make it unavailable for deteriorative reactions [21]. The oxygen scavenger market is expected to reach 2.41 billion dollars by 2022, at a compound annual growth rate (CAGR) of 5.1% from 2017 to 2022 [21]. The most commonly used oxygen scavenger is ferrous oxide, but other compounds include catechol, ascorbic acid, sulfites, photosensitive dyes, unsaturated hydrocarbons and enzymes such as glucose oxidase and catalase [22, 23].

Free Radical Scavengers

When UV light, oxygen or metals cannot be effectively removed from the packaging atmosphere or food product, oxidation reactions start to take place in the food product and its atmosphere. This results in the formation of free radicals, such as oxo, hydroxo or peroxo that initiate oxidative chain reactions [24]. Free radical scavengers react with these free radicals to convert them into more stable compounds that are not able to engage in further oxidative chain reactions. There is a multitude of available antioxidant compounds that are able to scavenge free radicals, ranging from the synthetic ones such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA) and tert -butylhydroquinone (TBHQ) to the natural ones such as natural compounds, plant and fruit extracts and essential oils from herbs and spices [2]. There are also some developments based on nanoparticles such as selenium (Se) nanoparticles, incorporated behind a layer of LDPE in a multilayer, using the adhesive as vehicle [25]. This new antioxidant packaging material has been recently approved by EFSA. The current tendency is to move from synthetic antioxidants towards the natural alternatives, to circumvent the safety issues with the synthetic ones, namely their potential harmfulness to humans [26]. Although natural antioxidants look promising and have already demonstrated their effectiveness as free radical scavengers [25, 27], usually they are required in large quantities in the packaging to attain the same antioxidant activity as the synthetic ones in the food product.

There is a tremendous amount of research on the development of new antioxidant packaging solutions; therefore, for the interest of this book chapter, only research from the last five years with proof-of-concept (assays in food systems) will be considered (Table 4). Regarding the polymeric matrices used for the packaging material, research has driven from plastic materials, either as a monolayer or multi-layer assembly, such as polyethylene (PE) or polyvinyl alcohol (PVA), to more environmental-friendly and sustainable materials as paper and board, bioplastics such as poly(lactic acid) and biopolymers such as the ones based on starch, proteins and chitosan. In these matrices, the most preferred antioxidants incorporated are of natural origin, such as natural extracts from fruits, green tea, olive leaf; food by-products such as polyvinylpolypyrrolidone washing solution; compounds from natural origin such as resveratrol, α-tocopherol and lycopene; and essential oils such as the ones from cinnamon and clove. All of these strategies were able to effectively inhibit lipid oxidation in mushrooms, beef, pork and poultry meat, fish, butter and oils, and processed foods such as chocolate peanuts and cereals. The decrease in food oxidation was measured through the determination of peroxide values (PV), thiobarbituric acid reactive substances (TBARS), formation of dienes and trienes, enzymatic assays as the tyrosinase assay, measurement of oxidation products such as metmyoglobin, hexanal and several ketones and aldehydes, and also degradation products of fatty acids.

Table 4Examples of antioxidant packaging strategies described over the last five years. AP-active packaging; NR-not mentioned.MatrixOrigin (Synthetic or Natural)Antioxidant CompoundDosagePackaging TypeMain FindingsRef.Molecularly imprinted hydrogelsNaturalFerrulic acid11 µmol g-1Cover film for canProtection of butter from oxidation evaluated by the TBARS test 50% decrease in TBARS values in AP packaged butter Oxidation was reduced by 25% after four weeks of butter storage[28]ChitosanNaturalKombucha tea1-3% (w/w polymer)Cover filmInhibition of lipid oxidation in minced beef patties after 6 days of storage evaluated by the TBARS test[29]Poly(lactic acid) (PLA) nanofibersNaturalα -Tocopherol5% (w/w polymer)Cover filmInhibition of lipid oxidation in raw beef evaluated by the TBARS test[30]PolyurethaneNatural and syntheticα-tocopherol and butylated hydroxytoluene (BHT)1-5% (w/w polymer)FilmPreclusion of soybean oil oxidation, evaluated by FT-IR, during 60 days at 60 ºC in the dark[31]LDPENaturalPolyvinylpolypyrrolidone washing solution (PVPP-WS) extract3-20% (w/w)Wrapping filmReduction of lipid oxidation in raw beef, with reductions in TBARS values higher than 60% at the 9th day of cold storage At the highest concentration applied (20%),[32]the onset of oxidation was delayed until the ninth dayLinear LDPENaturalResveratrol1% (w/w)BagsMaximum oxidation reduction of 34.7%, in raw beef evaluated by the TBARS test[33]Fish protein isolate (FPI)/fish skin gelatin (FSG)NaturalZinc oxide nanoparticles (ZnONP) Basil leaf essential oil (BEO)3% ZnONP (w/w, based on protein content) and 100% BEO (BEO) (w/w, based on protein content)Wrapping filmReduction of lipid oxidation in sea bass slices as evaluated by the PV and TBARS tests AP wrapped fish showed a 30-40% and 50-60% decrease in PV and TBARS, respectively[34]NM (plastic)NaturalGreen tea extractNMCover film and bagReduction of oxidation in chocolate peanuts and cereals Lower pyrazine and fatty acid oxidation Shelf-life extension from 9 to 18 months Decreased rancidity[35]Paraffin coated paperNaturalCinnamon essential oil100 g of cinnamon EO in paraffin emulsion " m-2"PadDecreased sliced mushrooms browning by decreasing tyrosinase activity No significant colour alterations (browning) after 9 days of storage[36]Cassava starchNaturalLycopene2-8% (w/w filmogenic solution)PouchesDuring storage, sunflower oil showed higher stability when stored in AP films with PV inside the limit for vegetable oil[37]Cassava starchNaturalLycopene2-8% (w/w filmogenic solution)PouchesNo significant differences in conjugate dienes and trienes formation between AP and control package[37]PVA-cyclodextrin-gelatin filmNaturalMango peel extract5% (v organic extract/v filmogenic solution)PouchesIrradiation together with AP decreased lipid oxidation in minced chicken meat as evaluated by the TBARS test[38]Soy protein isolateNaturalClove essential oil0.5% (v/v filmogenic solution)Wrapping filmDecreased tuna oxidation as observed by the decrease in TBARS values and aldehydes and ketones contents[39]ChitosanNaturalGinger and rosemary EO2% (v/v filmogenic solution)Wrapping filmsOxidation inhibition in minced poultry meat throughout storage time No significant differences in the results observed between the two EOs[40]Polyethylene filmNaturalOlive leaf extract2-15% (w/w adhesive)Cover filmReduction of oxidation as seen by a 25% decrease in TBARS values and the decrease in oxidative product bands in Raman spectroscopy[41]PVANaturalSeed cover extract of Zanthoxylum rhetsa1% (w/v film forming solution)PacketReduction of lipid peroxidation in ground chicken meat during the entire refrigerated storage period as observed by the lower TBARS values obtained Improvement in shelf-life up to 12 days at chilled condition[42]PENaturalGreen tea extractNMBagsImproves the shelf-life of minced pork meat duet to the reduction of Met myoglobin formation, an oxidation product of myoglobin Maintenance of meat redness for a longer period[43]LDPE-PET multilayerSyntheticSe nanoparticlesAdhesiveMultilayer filmImproves the shelf-life of cooked ham, potato chips, fresh meat, fresh chicken, vegetable salads and more[44]

Antimicrobial Packaging

Antimicrobials

The growth of microorganisms in food is one of the most important causes of food spoilage and an important cause of human disease. It has been estimated that about 25% of all foods produced globally are lost due to microbial spoilage [45]. Moreover, in the European Union, 4,786 foodborne outbreaks (including waterborne outbreaks) were reported in 2016 [12]. Microbiological food spoilage is caused by the growth of microorganisms which produce enzymes responsible for biochemical reactions that yield undesirable chemical compounds and cause changes in the original characteristics of the food product. These changes reduce the quality of the food product and decreased its shelf-life [46]. In addition, some of these microorganisms can also result in severe human illnesses (Fig. 1) if contaminated food is ingested. For instance, in 2016, EFSA (European Food Safety Authority) reported more than 400 death caused by the foodborne pathogens Campylobacter, Salmonella, Yersinia, Sigha toxin-producing E. coli and Listeria [12].

Foodborne microorganisms can come both from the natural flora of the product itself or from the contamination produced during harvesting, food processing, and distribution [47]. Therefore, there are a huge range of microorganisms that can be founded in food including bacteria, molds and yeasts. The type and load of predominant microorganisms in one specific food product depends mainly on their intrinsic characteristics (pH, water activity, composition, structure) and also on the storage conditions (temperature and atmosphere conditions mainly) [46]. For example, psychotropic Pseudomonas are commonly found in fresh meat stored aerobically at cold temperatures [48], while Brochothrix termosphacta grows mainly in fresh meat stored at cold temperatures and low levels of oxygen [49]. Considering the variability of existing foodborne microorganisms, the development of new strategies to reduce or inhibit the growth of microorganisms in food products represents a challenge even to developed countries. Moreover, the increasing consumer and producer demands for products with higher quality and longer shelf-life requires more efficient antimicrobial solutions [50].

Fig. (1)) 2016 data on zoonosis and foodborne outbreaks in Europe: a) reported number of confirmed human zoonosis; b) Distribution of strong-evidence foodborne outbreaks, by type of vehicle (excluding waterborne outbreaks). Source: The European Union summary report on trends and sources of zoonosis, zoonotic agents and food-borne outbreaks in 2016 [12].

In this context, the research dealing with the developments of new antimicrobial active packaging has increased as a viable alternative to reduce microbial growth in food [14, 51]. It is important to point out that each product type will need a specific antimicrobial packaging solution since its intrinsic characteristic and its microflora will be different. Given this, the type of antimicrobial substance and polymer selection will depend on the use of the material. For example, an active material intended for packaging refrigerated fresh meat stored aerobically should contain an antimicrobial active against Pseudomonas and a polymer impermeable to water. The number of active substances and polymers used in antimicrobial AP has increased over the last years, especially regarding those of natural origin that are the ones preferred by the consumer [14]. In addition, new technologies to control the release rate of the antimicrobial have also been incorporated into the material in order not only to prolong the release of the active compound, but also as a means to predict and yield more reproducible release rates [52].

Although the amount of research on the development of new antimicrobial AP technologies is huge, most of them do not include proof-of-concept tests with real food. This may be due on one hand, to the novelty of some of these technologies that are not fully developed and, on the other hand, to the difficulties encountered when applying the antimicrobial materials to real systems. It has been demonstrated that the efficiency in food systems is lower probably due to the interactions of the active compounds with the food matrix [53, 54], higher organic acid and trace metal contents, and greater availability of nutrients for microbial cell repair and survival [47]. Similar to antioxidant AP, in the case of antimicrobial AP, focus was given to relevant new antimicrobial packaging developments in last five years tested in food matrices (Table 5).

Concerning polymeric matrices, research includes classical synthetic polymers such as polyethylene or polyester used alone as monolayer material or combined to compose multilayer materials. Additionally, a high number of bio-based polymers have been investigated, namely poly(lactic acid) and those obtained from proteins, cellulose, polysaccharides (sodium alginate), chitosan or starch. Regarding the antimicrobial compounds evaluated, two main groups can de distinguished: synthetic nanoparticles (especially zinc oxide and silver) and antimicrobial compounds of natural origin. These include bacteriophages, antimicrobial peptides such as sakacin-A, ɛ-Poly-l-lysine, bacteriocins such as nisin, nanocellulose, essential oils (especially cinnamon essential oil) and natural extracts or compounds derived from plants such as pinosylvin or pomegranate peel extract. Chitosan is a particular case, since chitosan polymers have intrinsic antimicrobial properties. To assess the antimicrobial action of the active packaging, the reduction in Staphylococcus aureus, Salmonella typhimurium,Listeria monocytogenes, Escherichia coli, Campylobacter jejuni, total bacteria, moulds and yeast counts has been studied in fresh or read to eat meat (poultry, beef or turkey). Meat and meat products were the main food targets used probably due to their short shelf-life and high microbial load including both spoilage and pathogenic microorganisms. Other antimicrobial materials were also tested in fresh vegetables, bread, milk, fish or cheese. It is important to point out that the antimicrobial action achieved is not very high in all cases and, in some studies, the microbial reduction obtained is less than 2 log CFU.

Table 5Examples of antimicrobial packaging strategies described over the last five years. MIC- Minimal Inhibitory Concentration. CFU- Colony Forming Unit. PFU-Plaque Forming Unit.MatrixOrigin (Synthetic or Natural)Antioxidant Antimicrobial CompoundDosagePackaging TypeMain FindingsRef.Sodium alginate filmSyntheticZinc oxide nanoparticles3 mg mL-1Wrapping filmReduction of S. aureus and S. Typhimurium in poultry sausages at 8 °C (2 log CFU mL-1 in 24 h)[55]Low-density polyethyleneSyntheticCombinations of silver, copper oxide and zinc oxide nanoparticles1% (w/w polymer)Wrapping filmReduction of coliforms in ultra-filtrated cheese at 4 °C (4.21 log CFU g-1 after 4 weeks)[56]Poly(lactic acid) / poly-ethylenglicol blendSynthetic and naturalZinc oxide and silver/copper nanoparticles and cinnamon, garlic and clove oil1-4% (wt) nanoparticles 100% (w/w, based on PLA content) Essential oils 10-100% (w/w, based on PLA content)Wrapping filmFilms with cinnamon oil reduced L. monocytogenes and S. Typhimurium in cheese at 4 °C (4.47 log CFU g-1 and 3.17 log CFU g-1 respectively after 11 days)[57]Gelatin coated paperSyntheticCitric acid0.5 and 1.0% relative to the solution massWrapping filmReduction of total bacterial counts in beef at 4 °C (3 log g-1 after 4 days)[58]Zein filmNaturalZataria multiflora Boiss. essential oil10% (w/w dry zein power)BagsReduction of L. monocytogenes and E. coli in pasteurized cow's milk at 4 °C (0.99 log CFU mL-1 after 6 days)[59]Multilayer film made of polyester/aluminum/polyethyleneNaturalCinnamon and oregano essential oil and Benzoic acid18.5 and 10.2% (w/w dry adhesive)BagsFilms with cinnamon essential oil reduced E. coli O157:H7 (3 log CFU mL-1 after 24 h) and Saccharomyces cerevisiae (3 log CFU g-1 after 48h) in tomato puree[60]Cellulose / polypropylene absorbent padsNaturalPinosylvin0.08-0.8 mg cm-2 polymerAbsorbent padsGrowth reduction of C. jejuni in chicken exudates (56.99-99.99%) Growth reduction of total viable counts, total lactic acid bacteria and psychrotrophs in breast chicken fillets (10-74%)[61]Sodium caseinate filmNaturalPomegranate peel extract2 x MICCover filmReduction of total viable bacteria count (1.9 log CFU g-1) and Staphylococcus aureus (1.8 log CFU g-1) in ground beef at 4 °C after 9 days[62]Cellulose paperNaturalBacteriophage cocktails of Listeria monocytogenes and E. coli O104:H42.5x1010 PFU mL-1PadsReduction of Listeria monocytogenes in cantaloupes (1-3 log g-1) and turkey at 4 °C (0.8 log CFU cm-2)[63]Poly(lactic acid)NaturalSalmonella phage Felix O1 and Listeria phage A5111012 PFU mL-1Cover filmReduction of Salmonella sp. (0.8 log CFU cm-2 at 4 °C and 1.3 log CFU cm-2 at 10 ºC) andL. monocytogenes (6.3 log CFU cm-2 at 4 °C and 1.5 log CFU cm-2 at 10 ºC) in precooked sliced turkey breast[64]Low density polyethyleneNatural and syntheticNisin, chitosan, potassium sorbate and silver substituted zeolite2% (w/w polymer)Multilayer bagsReduction of mesophilic bacteria, total coliforms and total molds and yeasts in chicken drumsticks stored at 5 ºC after 6 days (0.28-1 log CFU g-1)[65]Poly (lactic acid) / sawdust particleNaturalBacteriocin 729319.54±2.87 μg cm-2PacketReduction of Gram-positive and Gram negative bacteria (2-5 log CFU cm-2) in chilled pangasius fish fillets[66]Polyethylene-coated paperNaturalSakacin-A0.44% (w/w dry weight of coating)Cover filmReduction of L. innocua in ready-to-eat thin-cut veal meat at 4 °C (1.5 log CFU g-1 after 48 h[67]StarchNaturalε-Poly-l-lysine1.6, 3.2 and 6.5 mg polylysine cm-2 of padCover biofilmFilms with 6.5 mg reduced A. parasiticus (2.73 log CFU g-1) and P. expansum (1.42 log CFU g-1) in bread[68]Chitosan-nanocellulose biocompositesNaturalChitosan and nanocellulose1% (w/0.6 v) chitosan 0.18% (w/w chitosan) nanocelluloseCover filmReduction of lactic acid bacteria in ground meat at 4 °C (1.3 log CFU g-1) and 25 °C (3.1 log CFU g-1) after 6 days[69]ChitosanNaturalEucalyptus globulus essential oil0.5-1.5% (v/v) of essential oilWrapping filmReduction of L. monocytogenes in sliced sausages at 23 °C (0.26-1.01 log CFU mL-1 after 3 days)[70]

Safety Assessment

According to European Regulations [4, 5], safety assessment of an active material implies both the safety assessment of the passive parts and the active compounds.

The passive parts of active and intelligent packaging systems refer to the materials or articles in which the active compounds are added or incorporated [71] and are subjected to pre-existing European and national food-contact material regulations such as Regulation (EU) No 10/2011 on plastic materials and articles intended to come into contact with food [72]. It is important to remark that in the case of the releasing systems, the amount of released active substance should not be calculated in the values of overall migration as the active substance is not part of the passive material (article 9(2) (EC) No 450/2009).

The active compounds, as defined in Regulation (EC) No 450/2009, are substances responsible for creating the active and/or intelligent function. These compounds must be subjected to the safety assessment carried out by the European Food Safety Authority before they are authorized for use. In the case of releasing systems, the released active substance should comply with relevant European Community provisions applicable to food as they can be incorporated into the food product. Moreover, active substances which are not in direct contact with the food or the environment surrounding the packaged food product and are separated from food by a functional barrier should not be evaluated if they comply with the requirement establish in article 5(2)(c) of Regulation (EC) No 450/2009.

In agreement with the European Regulations cited, the safety of a specific material implies that all the substances that can be released from the material to the food, either active compounds and substances from the passive parts, comply with the established limits of migration and levels of use in food. These limits have been established by the European Commission on the basis of EFSA scientific opinion on risk assessment made from data on estimated consumer exposure and results from toxicity studies. EFSA [73] indicated that migration data should give realistic account of migration into foodstuffs and mentioned three main approaches: modelling, simulation and direct measurement in foods (indicated when simulation is impossible or not reliable). Regulation (EU) No 10/2011 establishes the specific procedures for migration testing that should be applied to all food contact materials. According to this Regulation, migration from materials not yet in contact with food should be simulated. This means that materials should be exposed to one or several simulants (liquid o solid substances that mimic the behavior of food) during a selected time and temperature to simulate the real food exposure to the material [74]. The time, temperature and food simulant should be selected to simulate the worst case scenario in which the packaging is intended to be used. To a more detail review on active packaging migration please see [52]. After performing all the necessary migration tests, the identification of all compounds released by the new material has to be done, because either the active substances used or their impurities or their interaction products with the packaging material or with the food could contaminate the food with new toxic compounds. For instance, a migration study performed in several active packaging materials have demonstrated that impurities from one of the active agents, citral, were migrating from the packaging materials [75], showing the need for the analysis of compounds at trace levels. This identification procedure includes the qualitative and quantitative analysis of volatile and non-volatile compounds for which high resolution mass spectrometry is usually required.

Active Substances

As it has been explained in the previous section, the active substances that are incorporated into the active packaging material have to comply with the current legislation for approved substances in food. In many cases, they can have migration limits or maximum allowed levels in food and therefore, in this case, migration tests are mandatory.

Table 6 summarizes research carried out within the last five years on active (antioxidant and antimicrobial) packaging that carried out migration assays. As can be seen, it includes active materials based on different types of polymers since the material is a key factor in the migration process [52]. Specifically, synthetic polymers of polyethylene (PE), low and high density polyethylene (LDPE and HDPE), polyethylene terephthalate (PET), polyvinyl alcohol (PVA) or poly(butylene succinate) and other more environmental-friendly such as paper, bioplastics of poly(lactic acid) (PLA) or bio-based polymers of chitosan, proteins, starch, bacterial cellulose or peptides. Regarding the conditions used for the migration assays, in most cases, they only partially fulfilled with European Regulation (EU) No 10/2011 and used established simulants but different temperatures and times of exposure. It is important to clarify that the migration tests are conducted in some studies with the objective of evaluating the release behavior of the active compound from the polymeric matrix and not to assess the safety of the material.

Besides the conditions of the migration tests, the analytical methods used to detect and quantify the active substances in the simulants are essential to guarantee the compliance with the values of migration established, sometimes within the ppm and ppb level. As can be seen in Table 6, methods depend mainly on the type of substance analyzed. In the case of nanoparticles, the analytical technique mostly used is ICP (inductively coupled plasma) [76] combined with different detectors: MS (mass spectrometry), AES (atomic emission spectrophotometry) or OES (optical emission spectrophotometry) (Table 6). These techniques detect ultra-trace metals from metallic nanoparticles. To detect and measuring single, individual nanoparticles, SP (single particle) ICP-MS is used [77]. Atomic absorption spectroscopy is also used to detect metallic ions from nanoparticles [73]. Due to their volatility, gas chromatography is the most extensively analytical technique to detect and quantify the main compounds of essential oils [78]. To increase the sensitivity of this technique, a prior extraction technique such as headspace single-drop microextraction HS-SDME (single-drop microextraction) [79, 80] or solid phase microextraction SPME [60, 81] can be used to concentrate the sample. HPLC (high performance liquid chromatography) or UPLC (ultra-high performance liquid chromatography) coupled to different detectors as DAD (Photodiode Array Detection) [82] or MS [83] among others, occupies a leading position in the analysis of phenolic compounds [84] as thymol, quercetin or α-tocopherol (Table 6). Then, these techniques are used to quantify the migration of compounds of natural substances that are rich in phenolic compounds as essential oils such as cinnamon essential oil or natural extracts such as rosemary (Table 6).

Although less sensitive, spectrophotometric analysis is also employed with natural compounds that can absorb at different wavelengths. In table 6, this method is used to measure blueberry pomace, green tea extract and carvacrol. In some cases, compounds are previously subjected to chemical reactions to obtain colored compounds that can be measured using spectrophotometry. This is the case of epigallocatechin gallate that is measured after reaction with the Folin-Ciocalteu reagent and sulfur dioxide released by packaging material containing metabisulfite which is analyzed by using the p-rosaniline method (Table 6).

Up to date, there have been several strategies to mitigate the migration of active substances from packaging films, by providing a controlled-release of the active agent over time. These strategies include alterations in the polymer structure, polymer coating, polymer blends and composites. Over the past years, nanomaterials such as nanoclays, cellulose nanofibers and crystals, carbon nanotubes, technology has been applied to packaging materials either to be used as fillers, and thus alter polymer structure; or as encapsulating agents for the active compounds and thus control active agent release directly. For a more detailed review [52].

Food Contact Materials

Food contact materials (FCM) refer not only to the packaging materials, but to any material or article in direct or indirect contact with the food product. This definition also involves those materials used in food processing, production and distribution, as during any of the food chain processes, the materials could transfer substances to the food product, resulting in food chemical contamination. The European frame regulation establishes that no FCM should transfer to the food any substance capable of endangering human health, being this the main rule to follow. However, the design of an active packaging implies that some substances will be released from the material and arrive at the food, at levels that could be even higher than those established by the regulation 10/2011/EU of plastics in contact with food; therefore, it is important to say that these active substances have to be approved as food additives, as food ingredients or accepted as active ingredients to be incorporated into the polymers. In the case of active

Table 6Examples of active packaging materials developed in last five years reporting migration assays.CompoundPolymerMigration AssayAnalytical Techniques for Migrant DeterminationMigration ResultsSafety RemarksRef.ThymolLDPESimulants: ethanol 10% (v/v) and ethanol 95% (v/v) Temperature: 40 ºC Time: 7 days Method: immersionHPLC-DADPartition coefficients of 15-82% Diffusion coefficients of 7.5×10−13- 3.0×10−12An ADI of 0.03 mg kg-1 body weight (bw) day-1 is the agreed value stated within the Commission Review report for thymol (SANCO/10581/2013 rev 3) [85][82]PLASimulants: 15% and 95% ethanol Temperature: 30, 40, 50 and 60 ºC (ethanol 95%); 60, 65, 75 and 83 ºC (ethanol 15%) Time: 10 hours Method: immersionGC-MSincreased the temperature increases the release rate of thymol, with equilibrium being reached within 1.4 h 87-100% thymol release to 95% ethanol over the temperature range (30-60 ºC) Diffusion coefficients of 0.29-5.75x1012 m2 s-1[86]CarvacrolStarch/ PolyesterSimulants: ethanol 10%, ethanol 50%, isooctane and acetic acid 3% Temperature: 20 ºC Time: 160 h Method: Direct contactSpectrometry UV-visibleMigration of carvacrol was lower in ethanol 10% and acetic acid 3%. The migration of carvacrol was faster in ethanol 50%Carvacrol is considered a flavoring substance [87] according to European Regulation[88]CurcuminPVASimulant: 50% ethanol/water Temperatures: 25, 35 and 45 ºC Time: 10 hours Method: immersionSpectrophotometric detection at 279 nmAlmost 100% of migration after 1 h of incubationClinical trials have shown that curcumin is safe, even when consumed at a daily dosage of 12 g for 3 months