Preservatives and preservation approaches in beverages. Volume 15, The science of beverages 9780128166864, 012816686X, 9780128166857

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Preservatives and preservation approaches in beverages. Volume 15, The science of beverages
 9780128166864, 012816686X, 9780128166857

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P R E S E R VAT I V E S FOR THE BEVERAGE I N D U S T RY Volume 15: The Science of Beverages Edited by


An imprint of Elsevier

Woodhead Publishing is an imprint of Elsevier The Officers’ Mess Business Centre, Royston Road, Duxford, CB22 4QH, United Kingdom 50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States The Boulevard, Langford Lane, Kidlington, OX5 1GB, United Kingdom © 2019 Elsevier Inc. All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Details on how to seek permission, further information about the Publisher’s permissions policies and our arrangements with organizations such as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website: www. This book and the individual contributions contained in it are protected under copyright by the Publisher (other than as may be noted herein). Notices Knowledge and best practice in this field are constantly changing. As new research and experience broaden our understanding, changes in research methods, professional practices, or medical treatment may become necessary. Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any information, methods, compounds, or experiments described herein. In using such information or methods they should be mindful of their own safety and the safety of others, including parties for whom they have a professional responsibility. To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume any liability for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas contained in the material herein. Library of Congress Cataloging-in-Publication Data A catalog record for this book is available from the Library of Congress British Library Cataloguing-in-Publication Data A catalogue record for this book is available from the British Library ISBN 978-0-12-816685-7 For information on all Woodhead publications visit our website at

Publisher: Andre Gerhard Wolff Acquisition Editor: Patricia Osborn Editorial Project Manager: Vincent Gabrielle Production Project Manager: Sojan P. Pazhayattil Cover Designer: Matthew Limbert Typeset by SPi Global, India

CONTRIBUTORS C. Anandharamakrishnan  Computational Modeling and Nanoscale Processing Unit, Indian Institute of Food Processing Technology (IIFPT), Thanjavur, India T.R. Ayora-Talavera  Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Guadalajara, Mexico Emmanuel Bajyana Songa  University of Rwanda, College of Science and Technology, Biotechnology Unit, Kigali, Rwanda Uttam C. Banerjee  Department of Pharmaceutical Technology, National Institute of Pharmaceutical Education and Research, SAS Nagar, India Rama Bhadekar  Department of Microbial Biotechnology, Rajiv Gandhi Institute of IT and Biotechnology, Bharati Vidyapeeth University, Pune, India Jinal Bhola  Department of Microbial Biotechnology, Rajiv Gandhi Institute of IT and Biotechnology, Bharati Vidyapeeth University, Pune, India Carlos Ariel Cardona Alzate  Institute of Biotechnology and Agribusiness, National University of Colombia, Manizales, Colombia Ömer Utku Çopur  Uludag University, Faculty of Agriculture, Department of Food Engineering, Bursa, Turkey Erika Tayse da Cruz Almeida  Laboratory of Food Microbiology, Department of Nutrition, Health Sciences Center, Federal University of Paraíba, João Pessoa, Brazil Jossana Pereira de Sousa Guedes  Laboratory of Food Microbiology, Department of Nutrition, Health Sciences Center, Federal University of Paraíba, João Pessoa, Brazil Evandro Leite de Souza  Laboratory of Food Microbiology, Department of Nutrition, Health Sciences Center, Federal University of Paraíba, João Pessoa, Brazil V Devi Rajeswari  Department of Biomedical Sciences, School of Biosciences and Technology, VIT University, Vellore, India Bhavya Kavitha Dwarapureddi  Department of Environmental Studies, GITAM Institute of Science, GITAM University, Visakhapatnam, India E. Gastélum-Martínez  Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Guadalajara, Mexico


xii  Contributors

Gargi Ghoshal  Dr. S.S. Bhatnagar University Institute of Chemical Engineering and Technology, Panjab University, Chandigarh, India Kristen Griffin  Clinical Laboratory Technology Program, Saint Louis Community College, St. Louis, MO, United States Bige İncedayi  Uludag University, Faculty of Agriculture, Department of Food Engineering, Bursa, Turkey Aprajeeta Jha  Department of Agricultural and Food Engineering, Indian Institute of Technology Kharagpur, Kharagpur, India V.N. Kalpana  Department of Biomedical Sciences, School of Biosciences and Technology, VIT University, Vellore, India Manoj Kumar Karnena  Department of Environmental Studies, GITAM Institute of Science, GITAM University, Visakhapatnam, India Prabhjot Kaur  Dr. S.S. Bhatnagar University Institute of Chemical Engineering and Technology, Panjab University, Chandigarh, India Navneet Kaur  Department of Chemistry, Panjab University, Chandigarh, India Hanna Khouryieh  Food Processing and Technology Program, School of Engineering & Applied Sciences, Western Kentucky University, Bowling Green, KY, United States Mara Krempel  Department of Chemistry, Western Kentucky University, Bowling Green, KY, United States A. López-Malo  Department of Chemical and Food Engineering, University of the Americas Puebla, Puebla, Mexico A.C. Lorenzo-Leal  Department of Chemical and Food Engineering, University of the Americas Puebla, Puebla, Mexico François Lyumugabe  University of Rwanda, College of Science and Technology, Biotechnology Unit, Kigali, Rwanda E. Mani-López  Department of Chemical and Food Engineering, University of the Americas Puebla, Puebla, Mexico J.A. Moses  Computational Modeling and Nanoscale Processing Unit, Indian Institute of Food Processing Technology (IIFPT), Thanjavur, India Carlos Eduardo Orrego Alzate  Institute of Biotechnology and Agribusiness, National University of Colombia, Manizales, Colombia Sebastián Ospina-Corral  Institute of Biotechnology and Agribusiness, National University of Colombia, Manizales, Colombia Azime Özkan-Karabacak  Uludag University, Faculty of Agriculture, Department of Food Engineering, Bursa, Turkey N.A. Pacheco-López  Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Guadalajara, Mexico

Contributors  xiii

Pinki Pal  Department of Chemistry, Birla Institute of Technology, Mesra, Ranchi, India E. Palou  Department of Chemical and Food Engineering, University of the Americas Puebla, Puebla, Mexico Jay Prakash Pandey  Department of Chemistry, Birla Institute of Technology, Mesra, Ranchi, India Pushap Raj  Department of Chemistry, Indian Institute Technology Ropar, Rupnagar, India M.O. Ramírez-Sucre  Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Guadalajara, Mexico M.A. Ríos-Corripio  Department of Chemical and Food Engineering, University of the Americas Puebla, Puebla, Mexico I.M. Rodríguez-Buenfil  Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Guadalajara, Mexico M.A. Sánchez-Contreras  Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Guadalajara, Mexico Gautam Sen  Department of Chemistry, Birla Institute of Technology, Mesra, Ranchi, India Filipa V.M. Silva  Department of Chemical and Materials Engineering, University of Auckland, Auckland, New Zealand; School of Agronomy, University of Lisbon, Lisboa, Portugal Amanpreet Singh  Department of Chemistry, Indian Institute Technology Ropar, Rupnagar, India Narinder Singh  Department of Chemistry, Indian Institute Technology Ropar, Rupnagar, India Sanelle van Wyk  Department of Chemical and Materials Engineering, University of Auckland, Auckland, New Zealand Saritha Vara  Department of Environmental Studies, GITAM Institute of Science, GITAM University, Visakhapatnam, India

SERIES PREFACE Food and beverage industry accounts among the most developed sectors, being constantly changing. Even though a basic beverage industry could be found in every area of the globe, particular aspects in beverage production, processing, and consumption are identified in some geographic zones. An impressive progress has recently been observed in both traditional and modern beverage industries and these advances are leading beverages to a new era. Along with the cutting-edge technologies, developed to bring innovation and improve beverage industry, some other human-related changes also have a great impact on the development of such products. Emerging diseases with a high prevalence in the present, as well as a completely different lifestyle of the population in recent years have led to particular needs and preferences in terms of food and beverages. Advances in the production and processing of beverages have allowed for the development of personalized products to serve for a better health of overall population or for a particular class of individuals. Also, recent advances in the management of beverages offer the possibility to decrease any side effects associated with such an important industry, such as decreased pollution rates and improved recycling of all materials involved in beverage design and processing, while providing better quality products. Beverages engineering has emerged in such way that we are now able to obtain specifically designed content beverages, such as nutritive products for children, decreased sugar content juices, energy drinks, and beverages with additionally added health-promoting factors. However, with the immense development of beverage processing technologies and because of their wide versatility, numerous products with questionable quality and unknown health impact have been also produced. Such products, despite their damaging health effect, gained a great success in particular population groups (i.e., children) because of some attractive properties, such as taste, smell, and color. Nonetheless, engineering offered the possibility to obtain not only the innovative beverages but also packaging materials and contamination sensors useful in food and beverages quality and security sectors. Smart materials able to detect contamination or temperature differences which could impact food quality and even pose a hazardous situation for the consumer were recently developed and some are already utilized in packaging and food preservation.


xvi  Series Preface

This 20-volume series has emerged from the need to reveal the current situation in beverage industry and to highlight the progress of the last years, bringing together most recent technological innovations while discussing present and future trends. The series aims to increase awareness of the great variety of new tools developed for traditional and modern beverage products and also to discuss their potential health effects. All volumes are clearly illustrated and contain chapters contributed by highly reputed authors, working in the field of beverage science, engineering, or biotechnology. Manuscripts are designed to provide necessary basic information in order to understand specific processes and novel technologies presented within the thematic volumes. Volume 1, entitled Production and management of beverages, offers a recent perspective regarding the production of main types of alcoholic and nonalcoholic beverages. Current management approaches in traditional and industrial beverages are also dissected within this volume. In Volume 2, Processing and sustainability of beverages, novel information regarding the processing technologies and perspectives for a sustainable beverage industry are given. Third volume, entitled Engineering tools in beverage industry dissects the newest advances made in beverage engineering, highlighting cutting-edge tools and recently developed processes to obtain modern and improved beverages. Volume 4 presents updated information regarding Bottled and packaged waters. In this volume are discussed some wide interest problems, such as drinking water processing and security, contaminants, pollution and quality control of bottled waters, and advances made to obtain innovative water packaging. Volume 5, Fermented beverages, deals with the description of traditional and recent technologies utilized in the industry of fermented beverages, highlighting the high impact of such products on consumer health. Because of their great beneficial effects, fermented products still represent an important industrial and research domain. Volume 6 discusses recent progress in the industry of Nonalcoholic beverages. Teas and functional nonalcoholic beverages, as well as their impact on current beverage industry and traditional medicine are discussed. In Volume 7, entitled Alcoholic beverages, recent tools and technologies in the manufacturing of alcoholic drinks are presented. Updated information is given about traditional and industrial spirits production and examples of current technologies in wine and beer industry are dissected. Volume 8 deals with recent progress made in the field of Caffeinated and cocoa-based beverages. This volume presents the great variety of

Series Preface   xvii

such popular products and offers new information regarding recent technologies, safety, and quality aspects as well as their impact on health. Also, recent data regarding the molecular technologies and genetic aspects in coffee useful for the development of high-quality raw materials could be found here. In Volume 9, entitled Milk-based beverages, current status, developments, and consumers trends in milk-related products are discussed. Milk-based products represent an important industry and tools are constantly been developed to fit the versatile preferences of consumers and also nutritional and medical needs. Volume 10, Sports and energy drinks, deals with the recent advances and health impact of sports and energy beverages, which became a flourishing industry in the recent years. In Volume 11, main novelties in the field of Functional and medicinal beverages, as well as perspective of their use for future personalized medicine are given. Volume 12 gives an updated overview regarding Nutrients in beverages. Types, production, intake, and health impact of nutrients in various beverage formulations are dissected through this volume. In Volume 13, advances in the field of Natural beverages are provided, along with their great variety, impact on consumer health, and current and future beverage industry developments. Volume 14, Value-added Ingredients and enrichments of beverages, talks about a relatively recently developed field which is currently widely investigated, namely the food and beverage enrichments. Novel technologies of extraction and production of enrichments, their variety, as well as their impact on product quality and consumers effects are dissected here. Volume 15, Preservatives and preservation approaches in beverages, offers a wide perspective regarding conventional and innovative preservation methods in beverages, as well as main preservatives developed in recent years. In Volume 16, Trends in beverage packaging, the most recent advances in the design of beverage packaging and novel materials designed to promote the content quality and freshness are presented. Volume 17 is entitled Quality control in the beverage industry. In this volume are discussed the newest tools and approaches in quality monitoring and product development in order to obtain ­advanced beverages. Volume 18, Safety issues in beverage production, presents general aspects in safety control of beverages. Here, the readers can find not only the updated information regarding contaminants and risk factors in beverage production, but also novel tools for accurate detection and control.

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Volume 19, Biotechnological progress and beverage consumption, reveals novel tools used for advanced biotechnology in beverage industry production. Finally, Volume 20 entitled Nanoengineering in the beverage ­industry take the readers into the nanotechnology world, while highlighting important progress made in the field of nanosized materials science aiming to obtain tools for a future beverage industry. This 20-volume series is intended especially for researchers in the field of food and beverages, and also biotechnologists, industrial representatives interested in innovation, academic staff and students in food science, engineering, biology, and chemistry-related fields, pharmacology and medicine, and is a useful and updated resource for any reader interested to find the basics and recent innovations in the most investigated fields in beverage engineering.

Alexandru Mihai Grumezescu Alina Maria Holban

PREFACE A good number of reasons make beverages one of the most active functional foods and they include efficient storage for shelf-stable and refrigerated products, tremendous convenience in distribution and the effortlessness in meeting consumer demand. Preservation is a key element in the industry of beverages. Along with developed preservation methods, a high amount of preservatives have been investigated and applied for ensuring beverage stability and protect from any type of deterioration. Preservatives have been clustered in various groups depending on their source, synthesis method, structure, and role. Some chemical preservatives can taint the flavor of beverages, while others have particular bio-effects (such as antifungal or antibacterial agents). In recent years, the research tendency is to develop or identify novel preservatives with a very specific effect and to ensure a limited range of side effects of such products. The aim of this volume was to reveal and discuss recent findings made on the field of traditional and new preservation methods and preservatives, empathizing on their impact in food and beverage industry but also on the consumers. The volume contains 15 chapters prepared by outstanding authors from India, México, Brazil, New Zealand, Turkey, Colombia, United States, and Rwanda. The selected manuscripts are clearly illustrated and contain accessible information for a wide audience, especially food and beverage scientists, engineers, biotechnologists, biochemists, industrial companies, students and also any reader interested in learning about the most interesting and recent advances in beverage science. Chapter 1, Preservatives in Beverages: Perception and Needs, by V.N. Kalpana et al., focuses on enhancing awareness about the detrimental effects of artificial preservatives and it advocates natural preservatives for ensuring excellent safety and preservation of substances, high therapeutic efficacy, and enhanced general health. Chapter 2, Characteristics and uses of novel and conventional preservatives for fruit drinks and beverages, by E. Mani-López et al., aims to provide relevant information regarding the properties and uses of novel and conventional preservatives for fruit drinks and beverages; preservatives’ sensory acceptability, legal status, and commercial availability will also be reviewed. Chapter 3, Traditional bio-preservation in beverages: Fermented beverages, by Prabhjot Kaur et al., discusses about the engineering aspects used for the manufacturing of fermented alcoholic and nonalcoholic


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beverages from variant sources like cereals, fruits, milk, and from other sources as well as biotechnology of microbial cultures used for fermentation to obtain high yield and better quality with excellent source of nutrients to improve cognitive properties and to make functional beverage. Chapter  4, Nonconventional preservation techniques: Current trends and future prospects, by Rama Bhadekar et al., explains the intricate techniques and methods involved in the development of current trends and ongoing researches in the preservation of beverages that pave a gateway to modern manufacturing practices. Chapter 5, Emerging nonchemical potential antimicrobials for beverage preservation, by Evandro Leite de Souza et  al., focuses on the potential use of nonchemical substances or compounds considered candidates for use as antimicrobials in beverages, namely essential oils and their individual constituents, bacteriocins, enzymes, and chitosan. Information on the use of these antimicrobials in combined application with other technologies to preserve beverages, as well as their impacts on quality characteristics of these products will be also presented. Chapter  6, Natural preservatives for nonalcoholic beverages, by Saritha Vara et al., explores natural preservatives from traditional to the latest developed and future trends in their production and applications. Chapter  7, Nonthermal preservation of wine, by Sanelle van Wyk et al., reviews the application of the following emerging nonthermal technologies for wine production and preservation: high-­pressure processing (HPP), pulsed electric fields (PEF), ultrasound (US), high-pressure homogenization (HPH), low electric current (LEC), ultraviolet irradiation and filtration. In this chapter, the effect of these technologies on wine quality and microbial inactivation was explored and the technologies were compared. The commercial viability of using nonthermal technologies to reduce or eliminate the use of sulfur dioxide in the wine industry was also discussed. Chapter  8, Optimizing beverage pasteurization using computational fluid dynamics, by Aprajeeta et al., elaborates novel approaches considering model fluid and beer itself and the brewery industry which can successfully use to have a precise control over the pasteurization process and thereby contribute to quality assurance. The paper also presents the potential of computational fluid dynamics as an optimization tool for the beverage industry. Chapter 9, Process and impact of the addition of biocompounds on the development of pasteurized healthy juices, by Ramírez-Sucre M.O. et al., compiles the knowledge on the impact of processing and storage on the development of juices focused on its antioxidant capacity. Chapter 10, Preservation of beverage nutrients by high hydrostatic pressure, by Azime Özkan-Karabacak et  al., includes the effects of high hydrostatic pressure application on the nutritional parameters of beverages.

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Chapter 11, Prebiotics in beverages: From health impact to preservation, by Ospina-Corral Sebastián et al., contains a brief introduction about prebiotics and their benefits followed by the description of the prebiotics that are suitable for use in beverages, recent trends, and future perspectives of prebiotics in beverages. There is also an analysis of the current industrial production and the state of the art in the research of prebiotics in functional beverages, stability issues, the safety aspects related to prebiotics beverage consumption, and the examination of regulatory issues. Chapter 12, Processing techniques for mycotoxins—A balancing act of food safety and preservation, by Pinki Pal et al., evaluates the preservation and processing techniques and their calculated risks involved with respect to the uncertainty of the presence of mycotoxins. Chapter 13, Hydrocolloids as emulsifiers and stabilizers in beverage preservation, by Mara Krempel et al., provides an overview of the ingredients used in beverage emulsions, specifically the most significant protein and polysaccharide hydrocolloids, and discusses how they can be utilized to stabilize and protect flavors in beverage emulsions. Chapter 14, Detoxification and sensing of organophosphate-based pesticides and preservatives in beverages, by Amanpreet Singh et  al., describes biocompatible catalysts that specifically degrade organophosphate contamination in natural beverages. The major emphasis is given to recent advances in metal complex-based biocompatible catalysts that are well known for phosphate ester hydrolysis and oxime-based green catalysts as decontamination agents for organophosphates. Chapter 15, Traditional fermented alcoholic beverages of Rwanda (Ikigage, Urwagwa, and Kanyanga): Production and preservation, by François Lyumugabe et  al., aims to document the production processes of these traditional Rwandan alcoholic beverages and to highlight, where appropriate, the technological advances made and to suggest what could be done for improving the production process and preserving the traditional alcoholic beverages of Rwanda for the future generation.

Alexandru M. Grumezescu University Politehnica of Bucharest, Bucharest, Romania

Alina M. Holban Faculty of Biology, University of Bucharest, Bucharest, Romania



V.N. Kalpana, V. Devi Rajeswari Department of Biomedical Sciences, School of Biosciences and Technology, VIT University, Vellore, India

1.1 Introduction The most affordable way of enhancing the health and well-being of the maximum number of people living in this world is by providing proper nutrition. It has been estimated that more than seven billion people in this world are consuming some type of beverage every day in order to meet their daily nutritional requirements and the entire quantity comes from around tens of billions of servings on a daily basis. There are different kinds of beverages such as energy drinks, milk, soda, hard liquor, beer, wine, fruit juice, tea, and coffee available and you can even find them in the form of blending any ingredients seen in water. In fact, the life of every human being begins fully bathed in a sea of fluids. The fact of the matter is that human beings as fetuses drink prior to birth as well. Every individual consumes a beverage soon after he/she is born, primarily with breast milk, and this habit of taking water remains until death (Wilson and Temple, 2016). There is a drastic increase in the consumption of energy drinks among younger people even after knowing the fact that energy drinks have adverse effects (Pandey et al., 2014). The growth of the international beverage industry is extremely fast every year because of the launch of innovative products like herbal nutritional pills, antiaging water, fresh energy drinks, and vitamin fortified water as well. This rapid growth puts forward a good number of analytical challenges. The ever evolving and fresh needs for assessing traditional drinks such as bottled water, alcoholic beverages, milk drinks, fruit juices, and sodas aggravate these challenges in a serious way. A classic example is the process of contaminating milk with melamine and baby milk formula. The quality of the composition and safety of every beverage needs to be supervised to keep track of the product consistency, adulteration, and contamination and Preservatives for the Beverage Industry. © 2019 Elsevier Inc. All rights reserved.


2  Chapter 1  Preservatives in Beverages: Perception and Needs

proper monitoring is also needed to make sure that there is regulatory ­compliance from raw ingredients like fruit additives and water to the ultimate product. Beverages can be defined as any drinkable liquid other than water. They can be grouped into three categories: alcoholic (Gassara et al., 2015; Chagas et al., 2012), nonalcoholic (Ghorbel-Bellaaj et al., 2012; Rao et  al., 2011), and dairy-based beverages (Higueras et  al., 2015). Nonalcoholic beverages can be classified in terms of sugar content, as sugared (e.g., fruit juices, fruit nectar, and concentrated fruit juice) and nonsugared beverages (e.g., tea, coffee, and tisanes) (Fernandes et al., 2008). The dairy-based beverages differ from the others by the high content of protein and fat forming emulsions (e.g., milk, milkshakes, and milk juice drinks). Beverage industries are always challenged to meet the needs of consumers, namely the demand for safe, healthier, and functional products. Enhancing minimally processed or fresh products intake seems to be the tendency in the market. Thus, food technologists search for compounds from natural resources to be used in beverages, for example, to increase their shelf life, introducing natural preservatives (Nunes et al., 2016), developing active packaging (Ferreira et al., 2014), improving clarification procedures (Nualkaekul et  al., 2012), and developing novel functional beverages. The practice of adding food additives to maintain flavor or improve the look and taste has been in prevalence for centuries. One perfect example is the utilization of sulfur dioxide in wines. Another instance is the use of phosphoric acid in soft drinks, which is being done to provide them with a more intense flavor. This method results in slowing down the antioxidant properties of bacteria and mold. For improving the flavor and maximizing the shelf life, beverages are being loaded with different types of food additives. The amount of nutritious carbonated water available in a majority of the beverage products comes to around 90%. The sweetening process of water is done using sugar and high-fructose corn syrup (blend of dextrose and fructose) is also used to sweeten the water. The taste of the beverages is enhanced by the sweeteners while the food additives that work as nutrition enhancers assist in making beverages more functional and nutritious. In order to curtail the microorganism growth, beverages and foods are added with the processed beverage and food preservatives. There are synthetic as well as natural preservatives available and they improve the shelf life without making any serious changes in the nutritional value, form, texture, and taste (Bomgardner, 2014). Additionally, they also prevent the product from becoming toxic, thus making them edible for a longer period of time. People always show allegiance to healthy, green, and safe foods. Since the demand is for these types of foods, there is a trend in the market to grow the

Chapter 1  Preservatives in Beverages: Perception and Needs   3

intake of optimally processed and fresh food products, thereby eliminating the availability of chemical additives, which comprises food preservatives (Boye and Arcand, 2013).

1.2 Beverages 1.2.1  Beverage Consumption The modern food industry provides various types of beverages. On the other hand, water and breast milk constituted the diet of the early ancestors (Wolf et al., 2008). The consumption of fluid, primarily water, is inevitable for sustaining human life and it is indispensable for proper mental as well as physical function (Popkin et  al., 2010; Lieberman, 2007). Moreover, as shown in Fig. 1.1, some health problems are the aftereffects of water loss. When the human body loses 1%–2% of body weight due to fluid loss, it can be defined as mild dehydration (Kleiner, 1999) and this condition can lead to problems such as CHD (coronary heart disease), functional impairments, and moderate dehydration. Although water is a very critical aspect in the survival of human beings, only a handful of countries accept water as a source of nutrition (Manz et al., 2002; Sawka et  al., 2005; Popkin et  al., 2010) and approaches to the water needs calculations are vague. The restricting factor for a generalization of the advocated water consumption is the total body water and dehydration variability. The water needs are impacted by many factors including the body size (Popkin et  al., 2010), thirst mechanism (Forshee and Storey, 2003), gender, age, physical activity (Garriguet et al., 2008), climate (Sawka et al., 2005), food habits, and the level of respiration (Lieberman, 2007).

Fig. 1.1  Dehydration problems.

4  Chapter 1  Preservatives in Beverages: Perception and Needs

There is a need for creating better awareness about several factors that have an impact on the consumption levels of the beverage. A strong relation can be established between age and gender and total daily beverage intake and choices. For nutrition policy makers, a proper understanding of the differences in beverage consumption is very important. Past studies show that while drinking water is the prominent water source in all age groups’ diet (Garriguet, 2008; Garriguet, 2008; Bellisle et al., 2010; Barquera et al., 2008, 2010; Bello and Al-Hammad, 2006), variation can be seen in the intake of other beverages based on the age groups. Variations have been found in the beverage intake of adolescents and children with respect to age in several countries (Garriguet, 2008; Barquera et al., 2008; Bello and Al-Hammad, 2006). Children’s preference is for drinking milk while soft drinks are the preferred beverage for adolescents (Storey et  al., 2006), and adults also show great interest in taking alcoholic beverages and tea or coffee (Bellisle et  al., 2010). According to EFSA (European Food Safety Authority), children (2–3-year-old boys and girls) must consume 1.3 L/day. In other words, the adequate intake (AI) of 2–3-year-old children should be 1.3 L/day. When it comes to children aged between 4 and 8 years, the AI needs to be 1.6 and 2.1 L/day, respectively, for boys aged between 9 and 13. For girls aged between 9 and 13, the AI has to be 1.9 L/day. As far as adolescents or adult females (14 years and above) are concerned, they need to consume 2.0 L/day and adolescent or adult males (14 years and above) have to keep the AI at 2.5 L/day (Agostoni et al., 2010). As far as the past studies are concerned, there is a variation between 0.8 and 2.6 L/day existing in total relative beverage consumption among different age groups (Garriguet, 2008; Bellisle et al., 2010; Barquera et al., 2008; Bello and Al-Hammad, 2006). However, they are unable to get up to the advocated AI of fluids. Apart from this aspect, a decrease can be seen in the proportional availability of water in the average diet over a period of time because of the fact that there is variation in the consumption patterns of the individuals with respect to an array of beverages that comprise one or several of the ingredients like carbonation, nonnutritive sweeteners, artificial and natural flavorings, caffeine, and sugar (Daniels and Popkin, 2010).

1.2.2  Beverage Consumption and Nutrition Intake Beverage intake variations observed during the past few decades have influenced the overall consumption of nutrition products. Beverages with a reduced quantity of nutrition took the place of nutrient-filled beverages such as natural fruit juices or milk (Blum et  al., 2005; Duffey and Popkin, 2006; LaRowe et  al., 2007) and the

Chapter 1  Preservatives in Beverages: Perception and Needs   5

c­ onsumption of various nutrients is adversely impacted by the sugar intake. There has been a reduction in the consumption of several minerals like vitamins such as riboflavin, Vitamins K and A, iron, magnesium, and calcium (Libuda et al., 2009; Yu et al., 2016). Furthermore, it has been reported that there is a positive link between excess consumption of sugar-sweetened beverages and improper nutrient consumption, unhealthy eating habits, and risk of getting vulnerable to childhood obesity. According to a cross-sectional study, milk intake is linked with sufficient consumption of several nutrients like vitamin D and calcium. At the same time, there is a negative association between the overall quality of the diet and proper intake of several nutrients, and intake of completely natural fruit juice and sugar-sweetened beverages (Marshall et al., 2005). Proper intake of milk is necessary for calcium absorption. Calcium is the most vital component in the diet for developing bone health in children as well as adolescents. Anyhow, milk is getting substituted with sugar- sweetened beverages, which leads to a reduction in calcium consumption and can create negative effects on bone health. Apart from the consumption of calcium, adolescents’ and children’s diet quality and consumption of different nutrients get influenced by increased intake of sugar- sweetened beverages (Libuda et al., 2009; Linardakis et al., 2008; Marshall et al., 2005). Moreover, dietary patterns have a relation with beverage intake. Several individuals follow unhealthy dietary patterns, including the Western diet, and they also follow unhealthy beverage patterns such as the excess intake of sugar-sweetened beverages (Libuda et al., 2008; McGartland et al., 2003; Forshee et al., 2006; Sanchez-Villegas et al., 2009).

1.2.3 Energy Intake From Beverages Many studies have been conducted on the intake of beverages and most of them have concentrated on energy or sugar consumption from beverages that are sugar added. It is mainly due to the probable adverse health effects caused by excess intake of these beverages (Dennis et al., 2009). WHO advocates a diet where refined sugar is the source for 10% of the total energy generation (WHO, 2003); anyhow, several studies have observed an elevated intake of sugar added beverage (Lasater et al., 2011). Therefore, the absorption of energy from sugar has become higher. Sugar embedded beverages offer 6% of the total energy consumption in Germany and natural fruit juice and ­sugar-sweetened beverages contribute around 10.7% of the total energy in the United States (Wang et al., 2008). When it comes to the adolescents in Australia, the contribution of sugar containing soft drinks to the total energy consumption is 7.5% (Clifton et  al., 2011). The contribution of sugar added beverages to

6  Chapter 1  Preservatives in Beverages: Perception and Needs

total energy intake is 6.7% for children aged between 1 and 4 and it comes to around 9.4% for children aged between 5 and 11 (Barquera et  al., 2008). As far as adolescents are concerned, sugar containing beverages offer 13% of the total energy consumption. Furthermore, a lot of studies reveal that food consumption is not impacted by the sugar- sweetened beverage consumption. Food intake does not come down due to the reduced satiety of sugar added beverages (Wolf et al., 2008). Thus, it is observed that people who take beverages prior to or even during the meal consume the exact quantity of calories as people who take energy free beverages. High energy intake can cause higher vulnerability to overweight. Libuda et al. (2008) made use of the data of the DONALD study and they discovered no relation between the intake of sugar added beverage and body mass index (BMI) and also the body fat percentage. They established a positive linkage between the BMI of adolescent girls and the intake of sugar added beverage. There were no differences found between the BMI of excess sugar added beverage consumers, nonconsumers, and medium consumers, and this study also did not identify any connection between the intake of sugar added beverage and the BMI. According to the NHANES data, increased total energy consumption becomes a reality with elevated intake of soda, pure fruit juice, and milk; nevertheless, no relation between the BMI of children and intake of the sugar-sweetened beverages was established. Coppinger et  al. (2013) conducted a cross-sectional study in London and found that there was no relation between the intake of sugar added beverages and the BMI of children aged between 9 and 13. Nevertheless, several other cross-sectional and longitudinal research studies observed good connection between the sugar added drinks and overweight (Berkey et al., 2004; Novotny et al., 2004). As opposed to sugar added beverages, water consumption prior to or with the meal controls or lowers hunger. Consequently, the energy consumption also comes down. Increased quantity of whole milk consumption results in high energy consumption and increased intake of cholesterol and dietary fat (Ludwig et al., 2001). Thus, skimmed milk or low fat intake in place of whole milk is more advantageous.

1.2.4  Beverage Categories A beverage, also known as a drink, can be interpreted as a liquid especially made for meeting the fundamental basic needs, and it is a very vital part of societal culture. Based on different studies that are going to be mentioned in this thesis at later stages, beverages can be classified into four categories. They are sugar, nutrient-based, alcoholic, and noncaloric beverages like water.

Chapter 1  Preservatives in Beverages: Perception and Needs   7  Sugar-Sweetened or Added Beverages (Low Nutrient) The intake of sugar-added or sweetened drinks, also known as SSB, and conventional soft drinks has always been on the rise. An example of SSB is fruit drinks that contain more than 100% fruit juice from fruit flavored or concentrated drinks with extra amount of sugar; all types of colas and carbonated beverages can be categorized as regular soft drinks. The consumption of these types of drinks is very high among children (Wang et  al., 2008). Extra sugars encircle a wide range of sweeteners that may comprise high-fructose corn syrup (HFCS), maltose, dextrose, glucose, fructose, and sucrose, which is utilized as the most dominant sweetener in a majority of sugar-sweetened or added beverages (Sun and Empie, 2007). Of the drinks taken by the Americans in 2000, sugar added beverages were the principal energy source, which contributed up to 9.2% of the total energy consumption (Block, 2004). This figure is two times higher than the 3.9% recorded during the later parts of the 1970s. By contrast, the consumption of energy through milk deteriorated from 8.0% to 5.0%, and as far as other types of beverage intake (fruit juices, fruit drinks, tea/coffee, and alcohol) are concerned, only small changes were reported (Nielsen and Popkin, 2004). Studies reassert the adverse health influence involved with increased consumption of sugar (Ludwig, 2002; Tahmassebi et al., 2006), and they also provide ample support for substitution of sugar-sweetened drinks with reduced-fat milk and pure and natural fruit juices (Rampersaud et al., 2003).  Nutrient-Based Beverages Nutrient-oriented beverages like fruit juices or milk can be described as foods that you take “because of their capability to produce physiologic satiety, most probably from alterations in their fiber and protein contents” (Almiron-Roig and Drewnowski, 2003). There are well-established documents that substantiate the true role of milk in the case of human health (vanStaveren et al., 2008; Elwood et al., 2008). It is hard to find a better source of minerals (primarily calcium), carbohydrate, protein, good fat, and vitamins than milk. Moreover, milk offers protection against health complications and diseases such as osteoporosis, cancers, cardiovascular diseases, and hypertension (Huncharek et al., 2008; Opotowsky and Bilezikian, 2003). Contemporary studies in children as well as adolescents showed a specific beverage intake pattern that reflects increased sugar-­ sweetened beverage intake and reduced nutrient-oriented beverages (Ballew et al., 2000; Bowman, 2002; Phillips et al., 2004). According to Storey et al. (2006) and Hirsch et al. (2008), displacement similarities have been found out among the adults in America; in contrast, the

8  Chapter 1  Preservatives in Beverages: Perception and Needs

beverage consumption patterns among the adults in Canada have not yet been clearly comprehended, needing further examination). Over the past few years, 100% fruit juice intake has been on the rise (Hirsch et al., 2008), and it has replaced the consumption of raw fruit partially. Anyhow, studies have shown that the replacement quality was negatively impacted by the storage and packing methods for juices (Díaz-Juárez et  al., 2009). The intake of cranberries is primarily intended for securing protection against bladder infection and the consumption of citrus fruit juices is intended for their health benefits on arterial pressure and cancers.  Alcoholic Beverages In the United States, there has been an increase in the average alcohol intake (liquors, spirits, cocktails, wine, and beer) among people who are taking them moderately and for the last 500 years, there has been a decrease among people who can be termed as heavy drinkers in terms of alcohol consumption, with no reduction in the cumulative incidence of disorders due to alcohol use (Zhang et al., 2008). For the Americans, beer constitutes one of the ten best energy sources (Mukamal and Rimm, 2008). Controversies always revolve around the topic of health impacts due to moderate alcohol consumption; however, mortality and morbidity can always be linked with high intake of alcohol.  Noncaloric Beverages Tea or coffee without any extra cream or sugar, diet beverages that are sweetened using artificial sweeteners and water are typical examples of noncaloric beverages. These types of drinks do not add to the total energy consumption from beverages. There is well documented evidence available for the health benefits from sufficient water intake like protection against functional and metabolic problems. According to Popkin et al. (2005), clear association is there between increased water intake and eating patterns that are healthy, and this tendency is more prevalent and more obvious among older and highly educated adults. Only below one-fifth of the US citizens take artificially sweetened drinks or diet soft drinks (17.5%). Although aspartame’s dangerous side effects have been reported in animals (Soffritti et al., 2007), no proof is available that substantiate aspartame’s carcinogenic or neurotoxic effects in human beings (Magnuson et al., 2007). Tea and coffee consumption has been prevalent among adults for centuries and caffeine is an ingredient of these beverages. In order to assess the health impact of caffeine, several in-depth research studies have been performed. One of the reviews performed by Health Canada suggests that negative effects like cardiovascular issues, improper calcium balance, and general toxicity cannot be associated with moderate caffeine intake consumption, that is, intake of 400 mg/day or less) (Nawrot et al., 2003).

Chapter 1  Preservatives in Beverages: Perception and Needs   9

1.3 Preservatives FSSAI states that a Preservative refers to a substance that is added to food to make it highly resistant, retarding, or inhabitant to the acidification, fermentation, and other types of food decomposition processes (Seetaramaiah et al., 2011; Lab Manual 8, 2012). The shelf life of the foods can be enhanced by the preservatives and it is being done by safeguarding foods against the problems created by microorganisms. When it comes to using additives for food processing, 26 prominent additive categories are utilized and preservatives are certainly one of them. The evaluation process is done several times to make sure that they conform to the safety standards put forward by the EFSA (European Food Safety Authority and SCF (Scientific Committee on Food). Food preservation means the use of one of the several techniques employed to safeguard food from getting deteriorated. There are many techniques available like chemical additives addition, smoking, pasteurization, irradiation, drying and freeze drying, pickling, and canning. The significance of the preservation of food is increasing and it has become a very critical aspect of the food industry because only a restricted number of people consume foods that are produced on their own lands. Another important thing is that several consumers are relying on buying and eating out of season foods. The principles and goals of food preservation are displayed in Fig. 1.2.

Fig. 1.2  Objectives and principles of food preservation.

10  Chapter 1  Preservatives in Beverages: Perception and Needs

1.3.1 Importance of Preservatives A log history can be associated with the preservatives when it comes to safe use of food products. In fact, food preservation is one of the ancient technologies employed in food processing and the most prevalent methods were smoking, dehydration, and heating. A majority of the preservatives these days are really fungistatic in terms of their action. It implies that they are extremely effective in preventing the growth of organisms like yeast, molds, and fungi. Their effect on bacteria is minimal but excellent protection can be achieved by utilizing a blend of preservatives having antibacterial properties. Temperature, nutrient availability, availability or nonavailability of oxygen, pH, and water activity are the factors that make an impact on the microbial growth in a food product. The primary objective of preservatives is to ensure and maintain food safety for human intake, sustain the nutritional value, and maintain the overall quality (Chemat and Khan, 2011).

1.3.2 Categorization of Food Preservatives Depending on the source and action mechanism of food preservatives, their categorization can be done. This is given in Fig. 1.3. One of the most common classifications is the division into Class I and Class II. Under the category of Class I preservatives, edible ­vegetable oils, vinegar, spices, glucose, dextrose, sugar, and common salt can be included. There is no restriction on adding class  I preservatives in any of the foodstuffs. There are different types of Class II preservatives. The most common ones are benzoic acid, including salts; of that sulfurous acid including salts, Nitrites or nitrates, Food preservatives Based on

Mechanism of action




Fig. 1.3  Classification of food preservatives.



Chapter 1  Preservatives in Beverages: Perception and Needs   11

and/or potassium and sodium, they being present especially in foods such as pickled meat and ham. Other class 11 preservatives are sorbic acid and its calcium, potassium, and sodium salts, Sodium or calcium propionates, Lactic acid’s calcium, potassium, or sodium salts, propyl, methyl, or Nisinparahydroxysodium, and benzoates diacetate (Ortega-Rivas, 2012).

1.3.3 Types of Food Preservatives  Natural Food Preservatives Natural food preservatives are things that can be easily found in the kitchen among the everyday cooking ingredients. For a very long time, mankind has been using them. The most popular natural preservatives include onion, rosemary extract, vinegar, alcohol, sugar, salt, and many more. They can be described as conventional preservatives and people use them at home to make juices, jams, and pickles. For improving the shelf life of the food, natural preservatives are utilized in cooked and raw form and they have the ability to maintain taste, aroma, and the food itself for an extended period of time undamaged (Ahmed, 2013).  Artificial Food Preservatives Artificial preservatives refer to the group of chemical substances that are being sprayed outside of food or added to food. These types of substances are added to specific medications to slow the contamination by bacteria or other types of spoilage or even discoloration by problematic organisms. The Federal Government has classified the majority of preservatives as food additives and the definition of these additives by the FD&C (Federal Food, Drug, and Cosmetic Act) of 1938 is “any substance, the intended use of which results directly or indirectly, in its becoming a component or otherwise affecting the characteristics of food.” There is a classification of food preservatives in which they are being referred to as safe (GRAS) and the government recognizes the existing scientific agreement or concord on their safety, depending on their usage before 1958 or well-established scientific information available. They are regarded as the best and highly efficient option for a prolonged shelf life and for the preservation objectives, they are simply the best. Parabens, etc., Sorbates, Sulfites, Nitrites, and Benzoates are the most popular preservatives available (Sharma, 2015).  Antioxidant Food Preservatives Antioxidants can be described as compounds that get engaged in the process of preventing or retarding the spoilage of foods due to oxidative mechanisms. They take up the role of preservatives by

12  Chapter 1  Preservatives in Beverages: Perception and Needs

Fig. 1.4  Role of antioxidants.

showing resistance to the effects created by oxygen on food and offer immense benefits to human health. Food deterioration and rancidity are the aftereffects of oxidation and they result in the creation of odors and off-flavors, discoloration, taste change, and texture deterioration (Silva et al., 2016). The classification and examples of antioxidant food preservatives are illustrated in Fig. 1.4.  Antimicrobial Food Preservatives Bacteria, molds, and yeast growth in beverages and food are a serious concern and they can be prevented with the help of antimicrobial food preservatives. The most popular preservatives utilized in beverages are wine, quasidrug drinks, whereas those used in soft drinks are parabens, isobutyl, propyl, isopropyl, p-­ hydroxybenzoic acid ethyl, benzoic acid, sorbic acid, and vinegar (Ochiai et al., 2002). The primary application of benzoic acid is in acidic food products like fruit oriented drinks and soft drinks. It is mainly due to its activity in the pH range 2.5–4.0. For foods with increased pH, sorbic acid is utilized. The leading benefits of parabens are their stable response to extreme temperature levels and better efficiency at much elevated pH, ranging from 3 to 8. Proper legislation exists in the use of these types of preservatives and there are clear-cut guidelines for maximum allowed concentrations in different types of food. It is thus essential to find out their quantity in foods and to make sure that the amount of antimicrobial preservatives are within the allowed limits (Meyer et al., 2007).

Chapter 1  Preservatives in Beverages: Perception and Needs   13  Antienzymatic Preservatives The target of these types of preservatives is the enzymes available in the food that keeps on metabolizing after the harvesting process. One such example is the enzyme phenolase which becomes functional immediately after a potato or apple is cut and the exposed surface turns brown. The activity of the phenolase is slowed down by acids like ascorbic acid and citric acid and it is being done reducing the pH really low that is quite difficult for enzyme activity. Ethylenediaminetetraacetic acid (EDTA) is a metal-chelating agent which has the ability to eliminate or detach the metal cofactors that several enzymes require. The fungal and bacterial enzymes find it extremely difficult to thrive because of the functioning of Chelators (Glevitzky et al., 2009).

1.4  History of Preservation Preservation of food is a normal household activity. It is not at all a novel practice to preserve food as the history dates back to ancient periods. During those times, the need for preservation was not very high because people never focused on food storage with the same intensity as they do today. Hunting was done to get the required food. Gradually, people began living in small clusters or groups and with the increasing population, human beings shifted toward an agrarian lifestyle. Thus, they realized the requirement for food storage and preservation. Although human beings have been preserving food somehow or other till the era of ancient civilization, considerable changes and developments in food preservation started occurring from the 18th century. The 18th and 19th centuries witnessed revolutionary scientific developments which had a strong impact on the food preservation processes (Gould, 2012). The most popular preservation methods in the 18th century were cold storage, pickling, sugaring, curing, and drying and as the population as well as the consumption of food began to increase, the requirement for something more advanced became imminent. It was during this period that a lesser known and inadequately trained scientist called Nicolas Appert introduced a fresh method. His new preservation method advocated sealing food inside cans or bottles that are sterile after the cooking process. It is highly popular today with the name canning. After the Civil War, William Underwood introduced this process in the United States and John L. Mason developed a canning jar comprising a metal cap and rubber gasket in 1858 and these products were manufactured on a large scale (Thorne, 1986). There was an issue with taking the air out of the container that was later addressed by Louis Pasteur. According to him, food can also be spoiled by particles in the air. He conducted a lot of

14  Chapter 1  Preservatives in Beverages: Perception and Needs

experiments and research before designing a swan neck flask for removing air from the containers that are used for preservation. Pasteur managed to boil yeast soups in flasks that do not contain air with the help of this exclusive design and found out that there were no contaminants available in the flask. He found out that microbes could be killed and the formation of undesirable flavors would be prevented when foods are heated at elevated temperatures. Pasteurization is the name given to this process (Thorne, 1986). Clarence Birdseye was the person who introduced the freezing process and it was done at his home in Canada. One day, he ate fish and left remaining in ice for some period of time, he found that they stayed fresh and could not notice any change in quality or flavor. Immediately, he developed a method for freezing foodstuffs. He made the metal plates that were soaked in calcium chloride brine really cold. The storage of food was done in these brines. It was regarded as one of the most effective and healthy means of preserving food. The first Birdseye freezer was introduced into the market in 1930 in Massachusetts (Thorne, 1986).

1.5  Food Preservation Methods Preserving prepared and harvested food for future intake is an ancient practice that has been in prevalence for several centuries. Actually, it was an essentiality borne out of the real need to survive in a hostile environment and nonavailability of fresh food was a major concern in these types of environments. The practice of drying foods goes back to older times, when the drying process of vegetables and fruits was performed in an open stove or in the sun. The historical methods of preservation were salting, sugaring, drying, prickling, and cold storage, which were used to preserve fruits, vegetables, meat, and fish. The emergence of food science technology has led to several modern methods of food preservation which are listed below.

1.5.1 Canning Canning is a food preservation method that employs the process of heating and sealing the food in containers for the purpose of storage. This method annihilates autolytic enzymes and microorganisms and is a good option for preserving food to take out during times of need. The issue of botulism poisoning can be prevented with canning because the endosperm coating makes the botulism bacteria highly resilient to heat. Anyhow, canning can result in the loss of water-soluble nutrients because these nutrients may get lost in the liquid available in the can. Canning is a common practice is several industries such as milk and meat in Ghana.

Chapter 1  Preservatives in Beverages: Perception and Needs   15

1.5.2 Freezing The growth of microbes is prevented by freezing because of the reduced temperature and the lack of water. This process builds an atmosphere that inhibits bacterial growth. Bacteria can be present on the food, but they lie dormant and start the replicating process when thawing is done. The longevity of the frozen foods comes up to months and years. Freezing is most prevalent in preserving items such as meat and fish (Thorne, 1986).

1.5.3  High Heating Processing (Pasteurization) This inactivates autolytic enzymes and also leads to the destruction of microorganisms. However, it also leads to loss of heat-sensitive nutrients (Thorne, 1986). In Ghana, this method is employed by the fruit juice industry. Most of the fruit industries use pasteurization as a preservative method.

1.5.4 Dehydration The inspiration for the modern dehydration food processes is the age-old drying process that was performed for preserving food. The basic assumption is that the water content in food is reduced to a certain extent to prevent them from cultivating microbial growth. Water actually helps increase the rate of deterioration in food. Dehydration leads to longer storage, small size, and weight (Rahman, 2007).

1.5.5 Ionizing Irradiation For the purpose of eradicating food deterioration-causing organisms, the ionizing irradiation process involves the method of food being exposed to a specified quantity of radiation. Foods like spices undergo changes in taste on heating and this mechanism purifies foodstuffs like them. In such a situation, the chemical makeup can be retained by the foods even after exposure and this aspect avoids the possibility of the reproduction of the microorganisms and toxin creation. The preservation effect can also be increased with increased radiation (Ibarz, 2008).

1.5.6 Chemical Preservatives Specific chemical substances can be added to retard the microorganism growth in a foodstuff. This approach stops the growth of microbes without affecting the nutrients. The most common preservatives take up the role of antimicrobials or antioxidants. The oxidization process of the constituents in food is stopped by the antioxidants,

16  Chapter 1  Preservatives in Beverages: Perception and Needs

thereby preventing it from turning rancid. The antioxidants also prevent black spot development when there is oxygen exposure. One example is an opened cut apple. The fungi and bacteria growth is arrested or stopped by the antimicrobial preservatives. Anyhow, there should not be any harmful chemical substance available. A lot of chemical food preservatives are used in the food industry. A survey conducted in some of the local manufacturers of soft drinks in Kumasi showed that most of them use benzoic acid as a preservative. They claim that it is the most readily available preservative on the market and is also not expensive. In Ghana, there is permission from the Ghana Standards Authority to use nonalcoholic preservatives like sorbic acid, orthophosphoric acid, sulfur dioxide, and benzoic acid and its salts in the beverage industry (Davidson et al., 2013).

1.6  Preservatives Used in Nonalcoholic Beverages The dietary pattern of the people hailing from the developing countries is getting impacted by nonalcoholic beverages in a very serious way. During the dry season, these types of drinks are considered as refreshing or after meal drinks in both urban and rural areas. Preservatives, flavoring agents, sugar, and 90% of water constitute in these types of drinks. Fig.  1.5 shows some good examples of

Fig. 1.5  Classification of beverages.

Chapter 1  Preservatives in Beverages: Perception and Needs   17

­ onalcoholic beverages. Soft drinks create a congenial environment n for microbes because their water activity is very high. Another reason is that they contain a lot of vitamins and minerals. Soft drinks show strong resistance to bacteria and microbe growth because of their low pH levels due to carbonation, preservatives addition, and sugar content (Ashurst, 2016). The physical properties of soft drinks and preservatives (antimicrobial) decide the kind of chemical preservatives utilized in soft drinks. Several factors play an important role in determining whether or not add to preservatives and which one needs to be added for controlling the microorganism growth. They include storage conditions, packaging, vitamin presence, and high or low pH of the product. Benzoic acid, sulfur dioxide, and sorbic acid and its salts are the most prominent preservatives permitted and utilized in soft drinks. For several centuries, people are using sulfites and sulfur dioxide as antimicrobials, which can be described as highly efficient preservatives. Sulfur dioxide was used even during the Roman period. During those times, sulfur was subjected to burning and the exposure of the unfermented juice to the fumes was done for assisting the wine preservation process. In the 19th century, people added sulfites to lime juices and lemon casks and lime juices for conserving the fruit juice and to prevent diseases such as scurvy on the ships that were sailing on the ocean. Bound forms of sulfur dioxide are less active compared to free forms of sulfites. In addition to the antimicrobial properties, sulfites are utilized as antioxidants. They are also used as color stabilizers and antibrowning agents (Osuntogun and Aboaba, 2004). Unwanted growth of microorganisms like yeast in soft drinks is controlled by sulfites which also take up the role of antioxidants to control the browning reactions. Some people, primarily asthma patients, may become vulnerable to allergic reactions due to sulfites and sulfur dioxide. These types of allergic reactions happen frequently when the usage of sulfur dioxide gas is involved or wine is absorbed. In both these situations, the quantity is normally higher compared to the quantity seen in soft drinks. There are in-depth rules for the mandatory labeling of the additives utilized in a product, as laid down by the European legislation, which help the consumers make the right decisions and choices and thereby keeping away from the additives consumption (Altay et al., 2013). When it comes to curtailing the growth of bacteria, molds, and yeasts, Sorbates (sorbic acid and its salts) are the most efficient preservative. Several chemical and physical properties of soft drinks like storage length, storage temperature, packaging, processing, and availability of other additives and pH decide the antimicrobial effectiveness of the sorbate. Since better solubility can be associated with salts in comparison with the acid form, they have been most

18  Chapter 1  Preservatives in Beverages: Perception and Needs

c­ ommonly used. Excellent antimicrobial properties can be linked with sorbic acid but increased levels may create a negative effect on the product’s taste. When the soft drink is extremely acidic in nature, benzoates and Sorbates are frequently utilized in combination (Lino and Pena, 2010). In different types of berries like cloves, currants, plums, cinnamon, and cranberries, benzoic acid is a natural ingredient. This acid is extensively used for inhibiting the growth of microbes in several products such as emulsified sauces, jams, and nonalcoholic beverages. Better stability can be linked with the salt of the benzoate compared to the acid form and their better solubility in water makes benzoates the most effective option for the soft drinks industry. The use of sodium benzoate has a rich history of more than 100 years and for retaining taste and quality, it is extensively used by the food and beverage industry. Approval has been granted in using it in soft drinks by the European Union and several global regulatory bodies in countries such as Japan, Canada, Australia, and the United States also have given permission to use sodium benzoate. It is truly an efficient option against bacteria, yeasts, and molds. It is highly beneficial for using in soft drinks like juice and carbonated drinks because it functions admirably well b ­ etween pH levels of 2 and 4. The drink composition makes a serious impact on the suitability and efficiency for use (Tfouni and Toledo, 2002). When there is a combined existence of ascorbic acid (vitamin C) and sodium benzoate as ingredients in drinks, specific conditions lead to the formation of benzene. Benzene formation gets aggravated in beverages when storage is done at higher temperatures for a long period of time. The benzene formation levels and frequency that happened in the past have not been taken into consideration for creating a public health risk, but methods have been developed by the soft drink industry to stop or optimize its incidence. There has been some reduction in the use of benzoates in recent years and it is mainly because of the innovative processing techniques employed. Anyhow, it is still essential to make use of these preservatives in certain drinks for retaining their quality. Investigation was done on the passion fruit juice production and preservation for controlling the amount of spoilage and improving the juice shelf life with the help of chemical preservatives. Several preservatives such as citric acid, benzoic acid, sugar, and a blend of benzoic and citric acid (under room temperature) were used to perform the juice preservation under room temperature. The findings show that when 30% of benzoic acid was used as a preservative, the juice managed to retain its taste, aroma, and color for 30 days at least. It seems to be the most effective choice. The juice that was kept under preservative option, such as 4% sugar, completely deteriorated after 3 days. The option of 4% citric acid retained its qualities for seven days

Chapter 1  Preservatives in Beverages: Perception and Needs   19

and a few more days, but after this duration, the aroma began deteriorating. The blend of 4% citric acid and 3% benzoic acid retained the juice qualities for a period of 2–3 weeks. Estimation was taken on the alcoholic content and it was identified that the juice comprising sugar and citric acid has the most increased percentage of alcohol. The pH was also changed by the preservation so as to make the pathogen survival an impossible task at such a reduced pH environment (Akpan and Kovo, 2005).

1.7  Preservatives in Alcoholic Beverages The common feature of an alcoholic beverage is the presence of ethanol, which is generally called alcohol. Three classifications can be done with alcoholic beverages. They include wines, beer, and spirits. Nowadays, the brewing process of Indian beer is done at different locations in the country and it can be described as top-fermented. When it comes to flavors and smoothness, Indian rums have managed to gain a good reputation. There is tremendous demand for Indian alcoholic beverage products such as gin, rum, whiskey, brandy, beer, wine, and white wine (Fig. 1.5) in the international market. India ranks third when the market size of the alcoholic beverages in the world is taken into consideration (Buglass, 2011). The fermentation process of alcoholic beverages is done from the sugars available in grains, berries, fruits, and other ingredients like milk, honey, tubers, and plant saps. They might be distilled for reducing the actual watery liquid to a liquid that is equipped with increased alcoholic strength. Among the malt family of alcoholic beverages, beer is the most popular member and this family also comprises porter, stout, ale, and malt liquor. Hops, rice, corn, and malt are used to make it. The alcoholic content available in the beer ranges between 2% and 8%. For making wine, fruit juices like plums, berries, cherries, grapes, and apple cider are fermented. The beginning of wine making takes place with the fruit harvest and under strict temperature settings; the fruit juice is subjected to fermentation in big vats. When it is fully fermented, the filtering process of the mixture is performed before aging, and finally it is bottled. A total of 8%–12% alcohol availability is there in unfortified or natural grape wines. Good examples are Sauterne, Chianti, Burgundy, and Bordeaux. Brandy or alcohol is added to fortified wines, which have 18%–21% alcohol content. Examples are muscatel, port, and sherry. Distilled spirits creation starts with the mashes of fruits, grains, or other types of ingredients. The fermented liquid generated is subjected to heating until the vaporization of the flavorings and alcohol takes place and then, it is drawn off before cooling and condensing back into a liquid. Water is always left behind and it is

20  Chapter 1  Preservatives in Beverages: Perception and Needs

discarded. The concentrated liquid is known as distilled beverage and brandy, rum, vodka. Gin and whiskey come under this category. The alcoholic content in these beverages ranges between 40% and 50%, although lower or higher concentrations are identified. When alcoholic beverage is consumed, the gastrointestinal tract absorbs it in a speedy manner due to the fact that it does not come under any digestive process. That is why alcohol increases to elevated levels in the blood in a reasonably short period of time. The distribution of alcohol from the blood is done to all parts of the body and a pronounced effect is created on the brain by the alcohol on which it exercises a depressant action. A characteristic pattern of the brain functions occurs under the influence of alcohol. First of all, depression of complex brain actions like inhibitions acquired from childhood, self-criticism, and judgment occur and the control loss leads to an excitement and elated feeling during the early phases. Because of this aspect, alcohol is perceived to be a stimulant, which is an erroneous concept. When alcohol consumption increases, the alertness of the person who consumes it comes down in a gradual way. The environment awareness gets vague and dim and the deterioration of the muscular coordination occurs and facilitation of sleep becomes a reality (Jha et al., 2013).

1.8  Analytical Methods for the Determination of Preservatives in Beverages In order to determine the preservatives available in food, there are a lot of analytical methods used. In order to find out the quantity of benzoic and sorbic acids, spectrophotometric and titrimetric techniques are employed that make use of a thin layer chromatography with minimal sample pretreatment. For the assessment of the preservatives by GC-MS GC or HPLC (Lab manual 8), the introduction of ­solid-phase extraction was done recently as a pretreatment procedure. Other sophisticated analytical techniques like capillary electrophoresis (CE) were used that require dilution and filtration as a sample treatment. A novel extraction technique, called stirbarsorptive extraction (SBSE), was also introduced. It utilizes 50–300 μL polydimethylsiloxane (PDMS) coated with stir-bars for determining the amount of preservatives in beverages (Chen et al., 2010; Ochiai et al., 2002).

1.8.1 Determination of Benzoic Acid in Carbonated Beverages Benzoic acid (BA) is a commonly used antimicrobial preservative in food and beverages, especially in carbonated beverages, as it ­presents

Chapter 1  Preservatives in Beverages: Perception and Needs   21

its strongest antibacterial activity at pH 2.5–4.0. BA has inhibitory effects on the proliferation of bacteria and yeasts, a major cause of food spoilage. Although the addition of BA can extend the shelf life of drinks and prevent nutritional losses, excessive intake of BA may cause diarrhea, abdominal pain, and other symptoms and even interfere with the intermediate metabolic processes of the body. Thus, the maximum allowed concentrations of BA in every kind of food are restricted by legislation. According to the US Food and Drug Administration, the restriction of BA addition in general types of food is kept as 1000 mg kg−1. In contrast, the Chinese laws keep BA addition at 200 mg kg−1 as far as carbonated drinks are concerned. Different types of conventional methods, including thin layer chromatography, spectrophotometry (Lin and Huang, 2008), gas chromatography (GC) (Gonzalez et  al., 1998), and HPLC (Cao et al., 2010), have been introduced for BA identification. However, these methods are time-consuming with complicated pretreatments and require expensive instruments (Pylypiw and Grether, 2000). Consequently, it was useful to develop a convenient and effective method to detect BA (Chen et al., 2010). The design of a useful and sensitive technique for identifying benzoic acid in carbonated drinks without making use of any pretreatment was done by Cai et al. (2018). Coupling of silica gel thin-film microextraction (TFME) was done to find out SERS (Surface Enhanced Raman Scattering) and the authors made use of the fast identification of BA in carbonated drinks. Tiny pieces of silica gel substrate were placed in an uncomplicated device that is made at home for conducting the TFME process and then determination of the BA content was done with the help of SERS after dropping colloidal gold onto the substrate. They found that the obtained SERS signals of BA were strong and of high reproducibility. The method was implemented in a successful way for determining BA in carbonated drink samples as high consistency was linked with the results with those utilizing high performance liquid chromatography, indicating that it is a speedy method for the identification of BA in carbonated drinks.

1.8.2 Determination of Sodium Benzoate and Potassium Sorbate Preservatives in Beverages In order to find out the potassium sorbate and sodium benzoate that have been added as preservatives to various samples that are commercially available in the local markets of Syria like energy drinks, beverages, soft drinks, and tomato sauce in a rapid manner, a ­reversed-phase HPLC analysis was done without any pre-extraction. On a 5 μm Purospher STAR RP-18 column (25 cm × 4.6 mm), procedures like potassium sorbate and sodium benzoate separation and determination were carried out with the help of caffeine in the form of an internal

22  Chapter 1  Preservatives in Beverages: Perception and Needs

standard (0.04 mg/mL) and also with UV detection at 235 nm. 40 °C was the column temperature. As far as the mobile phase was concerned, a percentage (25:75) was found at a flow rate 1.2 mL min−1. 20 μL was the volume of injection. Good resolution was used for fully separating the two elution peaks. This method provided a detection of potassium sorbate and sodium benzoate in a direct way with great authenticity and precision for the results without any extraction (Antakli et al., 2010).

1.8.3  Quantitative Determination of Sorbic Acid and Sodium Benzoate in Citrus Juice With the help of anion exchange HPLC, the development of a procedure has been performed that has the ability to detect microgram quantities available in two popular food preservatives known as sorbic acid and sodium benzoate in citrus juices. The elution process of the compounds from the column was done using a simple buffer system and identified with the help of a variable wavelength spectrophotometer. Detection can be done in a simultaneous way with this method. The needed sample preparations are filtration and centrifugation and HPLC evaluation time is below 10 mm. Outcomes, reiterated by phosphorimetric analysis and ultraviolet absorption, are precise and re-creatable. Evaluation of the fortified juice samples in the common ranges records values within 5% of the created concentration (Valeria et al., 2007).

1.8.4 Determination of Ethanol in Alcoholic Beverages With the help of direct injection gas chromatography and megapore polar column (CPWax 58 CB, 30 m, 0.53 mm), a fast and simple method was designed for identifying the ethanol content in alcoholic drinks by Wang et al. (2003). After adding the right quantity of internal standard, acetonitrile solution, the injection process of ethanol in the sample was done in a direct way into GC for assessment. Because the limit quantization (LOQ) and sample preparation stayed at 0.5 μg/ mL, this method took less than 8 min to get the results. It can be easily implemented in the case of alcoholic beverages with differing alcohol contents. The precision, speed, and simple sample pretreatment procedure are the main benefits of this method and they can work as a routine analysis technique in place of existing methods.

1.8.5 Determination of Alcohol Content in Alcoholic Beverages The most extensively utilized recreational drug on a global basis is alcohol or ethanol and there are stringent laws that regulate

Chapter 1  Preservatives in Beverages: Perception and Needs   23

the production, sale, and consumption of these drugs. In alcoholic beverages like spirits, beers, and wine, the alcohol content approximately stands in between 3% and 50% v/v. There should be precision, accuracy, and reliability as far as the analytical techniques employed for deciding the alcohol content are concerned. A 45 MHz low-field benchtop NMR (nuclear magnetic resonance) spectrometer was used in this study to find out the quantity of ethanol in various types of alcoholic beverages. For accurately quantifying ethanol, analytical methods known as standard addition and internal standard were employed. Acetonitrile or acetic used was utilized as the internal standard for both methods to perform the alcohol content quantification process and it is being done with the help of the peak area that correlates with the methyl peaks of acetonitrile, acetic acid, and ethanol. The findings revealed that the internal standard method provided values of percent alcohol that are in complete unison with the suggested label as reiterated by running the samples in a 400 MHz high-field NMR spectrometer with the help of acetic acid as the internal standard. The utility of a benchtop NMR spectrometer is clearly shown by this study and this method can offer an alternative technique for evaluating percent alcohol in various types of alcoholic products (Saac-Lam, 2016).

1.9  Adverse Effects of Preservatives A tremendous increase in the number of people who show more allegiance to beverage drinks in comparison with fruit juices can be seen. It clearly results in a hike in the number of fresh side effects. The number of hospital admissions has been increasing on a daily basis and it is because of the excess intake of these types of beverages (Inetianbor et al., 2015). Fig. 1.6 shows the most common problems identified in people who consume these beverages in a regular manner.

1.10  Pros and Cons of Food Preservatives 1.10.1 Pros Food preservatives enhance the life span of foodstuffs, which suggests that food spoilage is well protected. It results in reduced wastage and enhanced supply. Since toxins are removed, especially in meats, foods acquire good quality and they become safer options to consume. Due to the increased supply, the price of the foods can be kept affordable. Preservatives also play their part in enriching food by adding the much needed daily nutrition needs. A typical example is vitamin C to orange juice. An improved visual appeal is also

24  Chapter 1  Preservatives in Beverages: Perception and Needs

Fig. 1.6  Preservatives in beverages—impact on human health.

g­ enerated with foods containing preservatives. One such example is how meat does not turn brown when sodium nitrites and nitrates assist in retaining the color. Flour after bleaching becomes white is another example. The natural color of the wheat is brown but packaging will provide the information like bleached flour. Emulsification refers to a process that permits two substances like water and oil that do not blend to be mixed when preservatives are utilized. It is clearly evident in salad dressings.

1.10.2 Cons Allergic reactions are a major con of the preservatives and asthma patients are highly vulnerable to this problem. Preservatives can also be associated with cancer. Sodium nitrite and nitrate are utilized for meat and they contain cancer-causing chemicals known as nitrosamines. Children may end up facing a lot of harmful effects because of the presence of the preservatives in foods. Major health problems that can be associated with preservatives are severe headaches, occurrence of attention disorders, and other types of behavioral changes.

Chapter 1  Preservatives in Beverages: Perception and Needs   25

1.11  Alternatives to Artificial Preservatives Artificial preservatives are mostly considered safe but many of them have negative potential and life-threatening side effects. Researchers also claim that there is a substantial link between the increased levels of nitrites in food and increased level of deaths from Parkinson’s disease, Alzheimer's disease, and Type 2 diabetes. Sweating, Headache, and redness of skin also occur after consumption of monosodium glutamate. Sulfite containing foods also cause asthma and allergic reactions (Anand and Sati, 2013). This aspect has managed to create more intense interest in formulating natural alternatives for preservatives to improve the food safety and quality. Natural substances or extracts obtained from plants, animals, and minerals serve as beneficial preservatives. Listed below are a few examples of alternatives of artificial preservatives. Algin: It is a compound extracted from seaweed. It is also used to make milkshakes and to thicken ice cream. It is also used to extend the shelf life of food products. Basil extracts: Derived from Ocimum basilicum. It is a useful antioxidant and antimicrobial agent. Carrageenan: A compound extracted from Irish Moss Chondrus crispus, a type of seaweed. It is used to make puddings, ice cream, and milkshakes. It also makes foods jell and stabilizes the food to keep the color and flavor even.

1.12 Conclusion The main objectives of the use of preservatives are to improve the food’s shelf life and retain the quality for a higher duration of time. Several studies show that chemicals are used as preservatives and they cause a good number of side effects. The intensity of the side effects caused by the preservatives ranges from mild to serious. Sometimes, things can reach up to a life-threatening situation. Although there are certain risks in the use of preservatives, its importance and contributions to the packaged food industry cannot be overlooked. A lot of research needs to be done to find out the natural and harmless preservatives. Natural preservatives always stand tall compared to artificial preservatives because they are highly nontoxic in nature. Moreover, natural options provide a broad array of health benefits. In order to obtain and maintain good health, people should opt for products containing natural preservatives and should read labels of eatables, cosmetics, and pharmaceuticals carefully. The food manufacturer should give special attention during their formulation for healthy preservatives as a combination of different preservatives has been known to improve not only the shelf life of the product but also enhance the quality and health benefits.

26  Chapter 1  Preservatives in Beverages: Perception and Needs

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30  Chapter 1  Preservatives in Beverages: Perception and Needs

Rao, L., Hayat, K., Lv, Y., Karangwa, E., Xia, S., Jia, C., Zhong, F., Zhang, X., 2011. Effect of ultrafiltration and fining adsorbents on the clarification of green tea. J. Food Eng. 102 (4), 321–326. Saac-Lam, M.F., 2016. Determination of alcohol content in alcoholic beverages using 45 MHz benchtop NMR spectrometer. Int. J. Spectrosc. 2016, 1–10. Sanchez-Villegas, A., Toledo, E., Bes-Rastrollo, M., Martin-Moreno, J.M., Tortosa, A., Martinez-Gonzalez, M.A., 2009. Association between dietary and beverage consumption patterns in the SUN (Seguimiento Universidad de Navarra) cohort study. Public Health Nutr. 12 (3), 351–358. Sawka, M.N., Cheuvront, S.N., Carter, R., 2005. Human water needs. Nutr. Rev. 63 (s1), S30–S39. Seetaramaiah, K., Smith, A.A., Murali, R., Manavalan, R., 2011. Preservatives in food products-review. Int. J. Pharm. Biol. Arch. 2, 583–599. Sharma, S., 2015. Food preservatives and their harmful effects. Int. J. Sci. Res. Publ. 5, 1–2. Silva, N.K.V.D., Sabino, L.B.D.S., Oliveira, L.S.D., Torres, L.B.D.V., Sousa, P.H.M.D., 2016. Effect of food additives on the antioxidant properties and microbiological quality of red guava juice. Rev. Ciên. Agron. 47 (1), 77–85. Soffritti, M., Belpoggi, F., Tibaldi, E., DegliEsposti, D., Lauriola, M., 2007. Life-span exposure to low doses of aspartame beginning during prenatal life increases cancer effects in rats. Environ. Health Perspect. 115 (9), 1293. Storey, M.L., Forshee, R.A., Anderson, P.A., 2006. Beverage consumption in the US population. J. Am. Diet. Assoc. 106 (12), 1992–2000. Sun, S.Z., Empie, M.W., 2007. Lack of findings for the association between obesity risk and usual sugar-sweetened beverage consumption in adults–a primary analysis of databases of CSFII-1989–1991, CSFII-1994–1998, NHANES III, and combined NHANES 1999–2002. Food Chem. Toxicol. 45 (8), 1523–1536. Tahmassebi, J., Duggal, M.S., Malik-Kotru, G., Curzon, M.E.J., 2006. Soft drinks and dental health: a review of the current literature. J. Dent. 34 (1), 2–11. Tfouni, S.A.V., Toledo, M.C.F., 2002. Determination of benzoic and sorbic acids in Brazilian food. Food Control 13 (2), 117–123. Thorne, S., 1986. The History of Food Preservation. Barnes and Noble Books. Valeria, A., Lozano, M.C., Jose, S.B., 2007. Maria simultaneous determination of sorbic and benzoic acid in commercial juices using the PLS-2 multivariate calibration method and validation by high performance liquid chromatography. Talanta 73, 282–286. vanStaveren, W.A., Steijns, J.M., de Groot, L.C., 2008. Dairy products as essential contributors of (micro-) nutrients in reference food patterns: an outline for elderly people. J. Am. Coll. Nutr. 27 (6), 747S–754S. Wang, M.L., Choong, Y.M., Su, N.W., Lee, M.H., 2003. A rapid method for determination of ethanol in alcoholic beverages using capillary gas chromatography. J. Food Drug Anal. 11 (2), 133–140. Wang, Y.C., Bleich, S.N., Gortmaker, S.L., 2008. Increasing caloric contribution from ­sugar-sweetened beverages and 100% fruit juices among US children and adolescents, 1988–2004. Pediatrics 121 (6), e1604–e1614. WHO, 2003. Diet, Nutrition and the Prevention of Chronic Diseases: Report of a Joint WH. Wilson, T., Temple, N.J., 2016. How beverages impact health and nutrition. In: Beverage Impacts on Health and Nutrition. Springer International Publishing, pp. 3–9. Wolf, A., Bray, G.A., Popkin, B.M., 2008. A short history of beverages and how our body treats them. Obes. Rev. 9 (2), 151–164. Yu, P., Chen, Y., Zhao, A., Bai, Y., Zheng, Y., Zhao, W., Zhang, Y., 2016. Consumption of sugar-sweetened beverages and its association with overweight among young children from China. Public Health Nutr. 19 (13), 2336–2346. Zhang, Y., Guo, X., Saitz, R., Levy, D., Sartini, E., Niu, J., Ellison, R.C., 2008. Secular trends in alcohol consumption over 50 years: The Framingham study. Am. J. Med. 121 (8), 695–701.



E. Mani-López, M.A. Ríos-Corripio, A.C. Lorenzo-Leal, E. Palou, A. López-Malo Department of Chemical and Food Engineering, University of the Americas Puebla, Puebla, Mexico

2.1  Fruit Beverages Fruit beverages are produced from raw materials or preserved semifinished products (like filtered-clarified juices, sieved puree, and concentrates, among others). Their consumption has increased worldwide in the last years due to different factors. One of these factors is their nutrimental content being generally low in minerals, proteins, and fat while rich in vitamins (A, C, and B group), moisture, fiber, antioxidants, and polyphenols; so they could, probably, help to manage dietary deficiencies. These beverages are also important because some of them, like fruit juices and nectars, are listed in the healthy eating dietary recommendations (Akusu et al., 2016; Chueca et al., 2016; Horváth-Kerkai and Stéger-Máté, 2012, Kalia and Parshad, 2015; Petruzzi et al., 2017). A big portion of the world’s fruit production, especially orange, apple, grapefruit, mandarin, lemon, pineapple, grape, pear, tomato, pomegranate, and cranberry is processed into juices, followed by fruit nectars from fruits such as mango, guava, peach, apricot, passion fruit, papaya, soursop (guanabana), strawberry, banana, and tamarind (Reyes-De-Corcuera et al., 2014). Processing adds an economic value to different raw materials, such as transforming fruits into other food products, such as juices, nectars, and musts/pomaces that could be stored and sold, reducing waste and minimizing losses that could occur to the fresh fruit (Curil et al., 2017). Over the years, there has been an increase in processing technology, product Preservatives for the Beverage Industry. © 2019 Elsevier Inc. All rights reserved.



formulation, equipment design, and production of fruit beverages, resulting in a great range of fruit drinks differing in composition, raw materials, nutrient content, sensory quality, and packaging; because of the industry development with the propose to find differentiated products that meet consumers demands. Generally, fruit drinks are classified per their content and/or composition, being the main difference among them the brand name (Horváth-Kerkai and Stéger-Máté, 2012).

2.1.1 Classification Mainly fruit beverages could be categorized into two groups: fruit juice and fruit drinks. The first one is defined as 100% juice with no extra ingredients added, and the second one is a beverage made from fruits and other ingredients (like sugar), this kind of drinks usually have only 10% of fruit juice (Leschewski et al., 2016). Related to fruit content, Horváth-Kerkai and Stéger-Máté (2012) indicated that there are three categories of fruit drinks: juices and fruit musts, fruit nectars, and soft drinks with fruit content. Fruit musts and juices are obtained by a mechanical process and thus have the taste, color, and aroma of the original fruit, they are consumed fresh immediately after production or preserved by heat treatments (such as pasteurization). Therefore, additives, different from fruit, sugars, and acids are not allowed in fruit musts and juices. At the same time, this category is divided into two subcategories: transparent or cloudy juices. The first subcategory refers to filtered juices and the second one to juices with colloids (like citrus juices) and probably fruit fibers. Nectars, in the other hand, are made from fruit juices diluted with sugar syrup or with sieved juices from single or blends of fruit juices (Horváth-Kerkai and Stéger-Máté, 2012). On the other hand, Fellows and Hampton (1992) classified fruit beverages as follows: (1) juices, as its name mentions it is a fruit juice without additives, (2) nectars that contain between 25% and 50% of fruit solids and must be drank immediately after opening, (3) squashes that contain 25% of fruit pulp with sugar syrup, usually having preservatives and are diluted in water, (4) cordials that are known to be crystal clear squashes, and (5) syrups that are clear concentrated juices that have high sugar contents; these drinks are mainly preserved by pasteurization, but they maintain their natural acidity and/or their high contents of sugar (Reyes-De-Corcuera et al., 2014). Another way to classify fruit beverages is related to their shelf life, such as fruit drinks that must be drank immediately after opening (these kind of beverages could not need any preservative if properly processed and packaged) and those that could be stored between uses, containing allowed preservatives (Fellows and Hampton, 1992).


2.1.2  Quality Factors Fruit beverages have high contents of carbohydrates, mainly easily metabolized dextroses (such as fructose and/or glucose), vitamins, water, and complex nitrogen sources. The content of organic nitrogen is usually low and they are generally presented as free amino acids. Vitamin content depends on the type of fruit and the processes to which the product is submitted; however, most of these beverages are fortified. Also, they could have high contents of organic acids that are influenced by pH (Akusu et al., 2016; Chueca et al., 2016; Reyes-DeCorcuera et al., 2014). Quality of fruit beverages is given by different sensory (flavor, color, aroma, appearance, and texture), nutrimental (vitamins, minerals, and dietary fiber contents), and antioxidant factors (α-carotene, β-­ carotene, β-cryptoxanthin, anthocyanins, and lycopene); being some parameters of their quality: pH, soluble solid, soluble solid to titratable acidity ratio (expressed as percentage of different acids to BX), color, cloud, vitamin C, and pulp contents (Reyes-De-Corcuera et al., 2014). The quality of fruit drinks also depends on the quality of their raw materials. Fruit ripeness is a crucial characteristic, because before of the optimal fruit ripe there are less sugar content and aromas, and when overriped, they could have less color compounds and reduced acidity (sometimes reflected in vitamin C content). For example, acidic fruit juices have desirable high contents of sugar and distinctive aromas when they are in their optimal ripe (Curil et al., 2017; HorváthKerkai and Stéger-Máté, 2012). Flavor, one of the most important and complex aspects of fruit drinks, depends on different properties like viscosity, nonvolatile and volatile compounds, and pulp content; it could be affected by treatment temperature and time. However, in some cases (like in apple juice), the changes on flavor caused by thermal treatments do not affect the acceptance of the product (Reyes-De-Corcuera et al., 2014). Another important aspect with regards to fruit beverages is color, which is visualized at first sight by consumers, and could be affected by enzymatic action or by anthocyanins, lycopene, and/or polyphenols losses (Danişman et al., 2015; Reyes-De-Corcuera et al., 2014). Finally, some fruits have high contents of insoluble plant components (such as protopectin, fibers, hemicellulose, cellulose, starch, and lipids) and colloid compounds (pectin, polyphenols, and proteins) that could cause turbidity and precipitation in their juices. Depending on the finished fruit drinks, insoluble components and colloid compounds should be entirely or partially eliminated by clarification or filtration, with the propose of improving sensory (color, taste, aroma, and flavor) attributes (Horváth-Kerkai and Stéger-Máté, 2012; ReyesDe-Corcuera et al., 2014).


2.1.3  Pathogenic and Spoilage Microorganisms of Interest Deterioration occurs in fruit beverages when there is a loss of nutrients, physicochemical changes, and/or microbial growth (Niir Project Consultancy Board, 2012). Consumers usually reject foods that present changes in appearance, smell, or taste; this phenomenon is known as food deterioration or spoilage. Spoilage primarily occurs because of the proliferation of natural microbiota; in fruit beverages also because they are in contact with air and environmental microorganisms during handling (Chueca et al., 2016; Petruzzi et al., 2017; Aneja et al., 2014b). Among the deteriorative microorganisms, yeasts are the most common group related to fruit drink deterioration; they can grow at low pHs, high sugar contents, and low water activities, being fruit drinks an ideal growing environment because of their high contents of carbohydrates, organic acids, and complex nitrogen sources (Chueca et al., 2016). This group of microorganisms could cause turbidity, flocculation, pellicles, clumping, and production of CO2 and alcohol; mainly due to metabolites of yeast activity in fruit drinks (Kregiel, 2015; Aneja et al., 2014b). Another important microbial group is the molds, which grow at almost the same conditions as yeast and could present mycelial mats and musty or stale off-flavors in fruit beverages. Bacteria, contrary to the previous groups, are more sensitive to low pHs, being lactic acid bacteria (LAB) the most common bacteria group present in fruit drinks, causing mainly off-flavors (Chueca et al., 2016; Kregiel, 2015; Aneja et al., 2014b). On the other hand, pathogenic microorganisms could also be present in fruit drinks, generally because of contaminated raw materials (especially fruits). Lately, foodborne outbreaks related to fruit beverages have increased; therefore, spoilage and pathogen bacteria could be indicators of low quality in these kinds of drinks. Some of the pathogenic and spoilage microorganism that have been found in selected fruit beverages are listed in Table 2.1 (Petruzzi et al., 2017; Aneja et al., 2014b).

2.1.4  Main Preservation Methods Among preservation methods that can be applied to fruit drinks, thermal treatments are the main ones since they have provided undeniable results for a long time. These treatments are based on heating the products until inactivation of spoilage and/or pathogenic microorganisms. Due to low acidity products (pH below to 4.5) vulnerability to microbial contamination and growth, thermal treatments and/ or adding of preservatives, became mandatory to produce stable shelf life products (Petruzzi et al., 2017; Reyes-De-Corcuera et al., 2014).


Table 2.1  Pathogenic and Spoilage Microorganisms Reported in Selected Fruit Beverages Microorganism

Fruit Beverage


Apple and orange juice

Alberice et al., 2012; Bevilacqua et al., 2013a; Pei et al., 2014 Khallaf-Allah et al., 2015 Aneja et al., 2014a Somavat et al., 2013 Aneja et al., 2014a Parker et al., 2010 Aguilar-Rosas et al., 2013 Aganovic et al., 2014 Aneja et al., 2014a Chueca et al., 2016 Parker et al., 2010 Aganovic et al., 2014; Char et al., 2010


Alicyclobacillus acidoterrestris Acetobacter spp. Bacillus coagulans Bacillus subtilis Clostridium sporogenes Lactobacillus brevis Lactobacillus plantarum Leuconostoc spp. Leuconostoc fallax Listeria innocua

Orange nectar Orange and carrot juice Tomato juice Orange and carrot juice Papaya nectar Apple juice Tomato juice Orange and carrot juice Apple juice Papaya nectar Apple, orange, and tomato juice


Bacillus cereus Escherichia coli

Orange and carrot juice Apple, prickly pear, mango, tomato, orange, and carrot juice

Shigella flexneri Listeria monocytogenes

Orange juice Mango, pineapple, orange, and carrot juice Acerola, cashew, apple, and mango nectar blend Carrot, orange, and watermelon juice

Salmonella spp.

Salmonella Typhimurium Staphylococcus aureus

Papaya, soursop, and guava nectar Apple and orange juice Carrot and orange juice

Aneja et al., 2014a Ait-Ouazzou et al., 2013; Aganovic et al., 2014; Aneja et al., 2014a; García-García et al., 2015; Luis-Villaroya et al., 2015 Dewanti-Hariyadi, 2014 Backialakshmi et al., 2015; Firouzabadi et al., 2014; Kamdem et al., 2010; Ngang et al., 2014 Da Silva et al., 2011 Danyluk et al., 2012; Sinchaipanit et al., 2013; Dewanti-Hariyadi, 2014 Gabriel et al., 2015; Parker et al., 2010 Dewanti-Hariyadi, 2014; Park and Kang, 2013 Aneja et al., 2014a; Sinchaipanit et al., 2013


Alternaria spp. Cladosporium spp. Colletotrichum spp. Fusarium spp. Geotrichum spp.

Orange and carrot juice Orange and carrot juice Orange and carrot juice Orange and carrot juice Orange and carrot juice

Aneja et al., 2014a Aneja et al., 2014a Aneja et al., 2014a Aneja et al., 2014a Aneja et al., 2014a Continued


Table 2.1  Pathogenic and Spoilage Microorganisms Reported in Selected Fruit Beverages—cont’d Microorganism

Fruit Beverage


Penicillium digitatum Pichia spp. Rhodotorula spp. Saccharomyces cerevisiae

Orange and carrot juice Orange and carrot juice Orange and carrot juice Prickly pear, tomato, orange and carrot juice Apple and orange juice blend

Aneja et al., 2014a Aneja et al., 2014a Aneja et al., 2014a Aganovic et al., 2014; Aneja et al., 2014a; García-García et al., 2015 Aganovic et al., 2014; Tyagi et al., 2014a


Aspergillus flavus Aspergillus terreus Aspergillus niger Candida krusei Candida parapsilosis Curvularia Penicillium islandicum

Orange and carrot juice Orange and carrot juice Orange, carrot and tomato juice Orange and carrot juice Orange and carrot juice Orange and carrot juice Orange and carrot juice

Aneja et al., 2014a Aneja et al., 2014a Aganovic et al., 2014; Aneja et al., 2014a Aneja et al., 2014a Aneja et al., 2014a Aneja et al., 2014a Aneja et al., 2014a

Usually, fruit beverages are preserved by pasteurization, having the target to reduce close to 5 log of the most resistant microorganism detected in the specific product. Pasteurization can be accomplished by different techniques, such as, high temperature-long time (HTLT); this thermal process uses temperatures between 90°C and 120°C for times around 1–2 min, and it is based in outside heat generation, which is transferred into food by mechanisms of convection or conduction. Another technique is the one called high temperature-short time (HTST), which ensures product safety, maintaining in some cases desirable bioactive compounds. This method uses temperatures ≥80°C and times ≤30 s. Other variations are: mild temperature-long time (MLTL) with temperatures below 80°C and times longer than 30 s, and mild temperature-short time (MTST) also with temperatures below 80°C and periods of ≤30 s (Chueca et al., 2016; Petruzzi et al., 2017). When fruit beverages are in contact with high temperatures, even for short periods of time, there could be undesirable changes in sensory and composition quality, decreasing fruit drinks benefits to health. Sometimes, fruit drinks such as, blended fruit juices, are pasteurized more than once because when juices and nectars are extracted they are submitted to pasteurization, and then again when they are blended (before packaging) causing even more damage in the


product qualities (Aganovic et al., 2016; Chueca et al., 2016; Petruzzi et al., 2017). At the same time, pasteurization can inactivate some enzymes (peroxidase, polyphenoloxidase, pectin esterase, and polygalacturonase) that cause undesired changes. Such is the case of polyphenoloxidase, which is the one responsible for browning and degradation of polyphenols and natural pigments in some fruit juices, causing losses of the antioxidant activity and discoloration (Aganovic et  al., 2016; Petruzzi et al., 2017; Reyes-De-Corcuera et al., 2014). Fruit drinks can also be preserved by aseptic packaging, being another method that utilizes high temperatures; this method consists in sterilizing and processing the package and the fluid independently and then, under aseptic conditions, hermetically seals them when brought together. However, like pasteurization, these treatments could also cause undesirable effects leading to quality and freshness reduction, reflected in flavor, color, texture, appearance, nutrient, and pigment losses. Therefore, some no-thermal methods such as ultrahigh pressures, electric pulses, UV, and/or ultrasound, have been proposed in different studies (Aguilar-Rosas et al., 2013; Carbonell-Capella et al., 2017; Horváth-Kerkai and Stéger-Máté, 2012; Petruzzi et  al., 2017; Pillai and Shayanfar, 2015; Shah et al., 2016). On the other hand, chemical preservatives are also utilized for preserving fruit beverages, being the most common ones sodium benzoate and potassium sorbate. The type of chemical preservative to be utilized in fruit drinks depends on the selected properties (physical and chemical) of the beverage as well as of the preservative. Product pH, vitamin content, packaging, and conditions of storage also may influence the choice of the additive. However, consumers demand for more natural, fruit products has increased over the years (Kregiel, 2015; Rupasinghe and Yu, 2012). To minimize damages, as degradation of nutritional and fresh characteristics of fruit drinks, it is recommended refrigeration temperatures during storage and transportation (Aganovic et al., 2016; Chueca et al., 2016; Petruzzi et al., 2017). The preservation of quality factors is a key goal of the fruit beverage industry; therefore, the importance of maintaining equilibrium between safety and nutritional quality of raw materials (Petruzzi et al., 2017).

2.2 Preservatives Foods and beverages often contain different types of food additives, among which preservatives play an important role (PetanovskaIlievska et  al., 2017); these are included in one of the 26 major additive categories that are utilized in foods (Kregiel, 2015). Before the advent of preservatives, food was placed in containers such as


clay jars to preserve them from spoiling. Food storage can be traced back to every ancient civilization such as Egyptian, Greek, Roman, Sumerian, and Chinese (Anand and Sati, 2013). Preservatives are defined as substances that are added to products such as food or beverages to prevent, stop, and/or delay any food deterioration due to microbial growth. An ideal food preservative remains effective until the product is consumed (Gbonjubola and Josiah, 2012). The principal properties of preservatives include the following: retard or reduce the growth of undesirable microorganisms, fungi, and bacteria, do not affect food texture or taste, be safe for human consumption, and extend food shelf life (lengthen the time before a food product begins to spoil). Shelf life is determined by rates of growth of spoilage microorganisms and chemical degradation of food components (Akinwande et al., 2012). Fruit juices are important commodities in the global market, providing vast possibilities for new value-added products to meet consumer demand for convenience, nutrition, and health. Fruit juices are spoiled primarily due to proliferation of acid tolerant and osmophilic microbiota. There is also risk of food-borne microbial infections or intoxications, which may be associated with the consumption of fruit juices. In order to reduce the incidence of outbreaks, fruit juices are preserved by various techniques (Aneja et al., 2014b). Traditionally, the stability of fruit drinks and beverage has been achieved by thermal processing. However, thermal processing tends to reduce the product quality and freshness; therefore, preservatives are a good option because these products display several advantages such as retention of sensory qualities and nutritional values over traditional thermal processing (Rupasinghe and Yu, 2012). Food preservatives may be classified as natural and conventional, which play a very important role in the beverage industry, citric acid is a good example of a naturally occurring preservative, sodium benzoate and potassium sorbate are representatives of the second type. The choice of appropriate preservatives for fruit drinks and beverages should take into consideration specific product requirements, the type of spoilage organisms associated with it, the product pH, the intended shelf life, and the mode of application. The pH and nutritional parameters are among the most decisive factors in choosing a preservative. In general, preservatives are only effective when the initial microbial contamination level is low. Most microbiological problems arise because of poor quality raw materials and poor process hygiene which lead to overcoming of the preservation system applied during manufacture by the spoilage organisms (Riikka et al., 2011). Good hygienic practices are essential to guarantee the quality of the products (Sospedra et al., 2012).


2.2.1  Conventional Preservatives Conventional preservatives are chemical substances that stop the growth and activities of the microorganisms and help to preserve foods for a longer time without affecting their natural characteristics; they include antimicrobial agents and antioxidants (Fig. 2.1). Antimicrobial agents are used to prevent or inhibit spoilage caused by molds, yeasts, and bacteria (Dhaka et al., 2016). Some antimicrobial agents are benzoates, nitrites, calcium propionate, and sorbates. Antioxidants are agents that are utilized to prevent the oxidation caused in the food material. Selected antioxidants are butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), formaldehyde, and some organic acids (Seetaramaiah et  al., 2011). The majority of the preservatives that are commonly utilized today are conventional rather than natural (Anand and Sati, 2013) and have been utilized by the food industry for decades. During processing and storage of fruit drinks and beverages, they can suffer microbial contamination; thus, shelf life of beverages can be extended by the addition of conventional preservatives that are applied to improve their microbiological stability.  Sulfur Compounds and Benzoic and Sorbic Acids and Their Salts There are two types of packaged fruit drinks, those that are drank straight after opening and those that are consumed in portions, so

Fig. 2.1  Conventional preservatives commonly utilized in fruit drinks and beverages.


they must be stored between drinking times. The former groups should not require any preservative if they are properly processed and packaged. However, the latter group must contain a certain amount of permitted preservatives to have a long shelf life after opening. As soon as the juice is expressed from the fruit, it starts to deteriorate, both as a result of chemical activity (enzymatic action) and microbial spoilage. Chemical preservatives often supplement other types of preservation methods, to ensure an economical, safe, and flavorful product for months or even years after preparation (Yadav et al., 2014). Some research has revealed that the shelf life of fruit drinks without any preservative was 3.7 >6.0

nr nr nr nr nr nr

500 500 400 500 400 500

300 300 20 300 20 300

8.0 8.0 >3.4 8.0 >4.2 8.0



Brettanomyces bruxellensis Brettanomyces bruxellensis Brettanomyces bruxellensis

Saccharomyces cerevisiae Saccharomyces cerevisiae Saccharomyces cerevisiae Saccharomyces cerevisiae Saccharomyces ludwigii Enological yeast

Red Cabernet Sauvignon Red and white Red Dolcetto Syrah Red SO2-free Cabernet Merlot Red Syrah Red Cabernet Sauvignon Red Pinot Noir Rosé White Chardonnay Red and white nr White nr Rosé Liquor white Sauterne

Puig et al. (2003) Tonello et al. (1998) Tonello et al. (1996)

Lonvaud-Funel et al. (1994)


Lactobacillus plantarum Oenococcus oeni Pediococcus damnosus Acetobacter aceti Acetobacter aceti Acetobacter pasteurianus a

Red and white Red and white Red Red and white Red Red and white

HPP was carried out at room temperature, maintaining nonthermal conditions; nr, not reported.

Puig et al. (2003) Tonello et al. (1996) Puig et al. (2003) Tonello et al. (1996) Puig et al. (2003)

Chapter 7  Nonthermal Preservation of Wine   217

plantarum, Acetobacter aceti, Acetobacter pasteurianus, and Oenococcus oeni in red and white wine, resulting in 8.0 log reductions for all four bacteria. Tonello et al. (1996) found that 400 MPa applied for only 20 s resulted in >4.2 log reductions of A. aceti in wine. Tonello et al. (1996) also looked at the inactivation of Pediococcus damnosus in wine using 400 MPa for 20 s, which led to more than 3.4 log reductions. As evident in this section, HPP has the ability to successfully inactivate wine spoilage yeasts and bacteria using treatment pressures and times that are feasible for use in the wine industry.

7.4  Pulsed Electric Fields 7.4.1 Overview PEF is a promising alternative for wine preservation because it is a nonthermal technology, operated in continuous mode. Depending on the application, it can have a relatively low energy consumption compared to other food preservation technologies (Sulaiman et  al., 2017a). Since PEF is a continuous process, it can easily be integrated into existing industrial processes (Delsart et  al., 2015). In operation, short microsecond pulses of high electric field strength are applied to pumpable beverages flowing between two electrodes (Milani et  al., 2015). The shape of the applied pulses is either exponential, where the voltage rises quickly to its maximum before decaying slowly to zero, or square where it remains constant for a few microseconds (Puértolas et al., 2009; Huang et al., 2013; Milani et al., 2015). A typical PEF system is based on a high-voltage pulse generator along with a treatment chamber, suitable fluid handling system as well as monitoring and controlling devices (Milani et al., 2015). PEF is able to inactivate spoilage microorganisms, often without any significant effect on beverage quality (Puértolas et al., 2009; Milani et al., 2015). Microbial inactivation is thought to occur due to the formation of a potential difference across the microbial cell membranes (Fig.  7.2). This in turn causes the permeabilization or electroporation of the cell membrane, leading to the loss of intracellular fluids (Delsart et  al., 2015). The electric field strength required for the inactivation depends on the size and shape of the cell as well as the composition of its membrane. As the size of the cell increases, it becomes more sensitive to the applied electric field pulses. Therefore, yeasts are more sensitive than bacteria (Milani et al., 2015). Also, rod-shaped cells required five times stronger electric fields than spherical cells of similar dimensions (Delsart et al., 2015). Most authors have reported a temperature increase associated with the increasing treatment of electric field strength or energy (Milani et  al., 2015). This increase in temperature also increases the rate of

218  Chapter 7  Nonthermal Preservation of Wine

Fig. 7.2  Diagram showing the pulsed electric field (PEF) inactivation of wine spoilage microorganisms (T refers to temperature).

inactivation by improving the cell membrane fluidity and mass transfer from the cells (Delsart et al., 2015). As this temperature increase is not desirable for wine, proper processing conditions must be selected, namely flow rate, initial wine temperature, and electric field intensity. Examples of PEF processed beverages include grape-, strawberry-, orange-, and apple juices, milk, soup, liquid egg, and beer (GardeCerdán et  al., 2008; Puértolas et  al., 2010b; Huang et  al., 2013; Abca and Evrendilek, 2014; Milani et al., 2015). PEF can also be operated or designed as a batch process, thus cured fish and meat products have also been processed (Puértolas et al., 2012). One of the main reasons why PEF has not been commercially implemented includes the need for high-voltage electric pulses, which pose a significant safety risk in commercial scale operations.

7.4.2  Impact on Wine Quality The effect of PEF treatment on the quality of finished wine is summarized in Table 7.3. Abca and Evrendilek (2014) found that subjecting red wine to 3 μs bipolar PEF pulses at 31 kV/cm (T ≤ 40°C), resulted in no changes to the sensory quality and the treatment had no effect

Chapter 7  Nonthermal Preservation of Wine   219

Table 7.3  Effect of PEF on Wine Quality Parameters Medium

Processing Conditions

Processing Time (μs)

Red wine

31 kV/cm, 3 μs square bipolar pulse, 40 mL/ min flow rate, T ≤ 40°C

Red wine

31 kV/cm, 1 Hz, 100 pulses, T  L*fresh juice; a*treated juice > a*fresh juice). CDs CDs/Hesp Moreover, color differences were clearly observed in the samples with longer heat treatment applied; however, only the juices added with the encapsulated hesperidin preserved the pleasant taste and smell (data not shown).


9.3.2  Product at Pilot Plant Level

3.85 3.80 3.75









Ascorbic acid (g/L)

0.25 0.20 0.15 0.10 0.05 0.00

The thermal process design for citric added with thermolabile antioxidants beverages becomes even more complex with all the size implications at pilot plant level. In a pilot-scale study, polyethylene terephthalate (PET) bottles were used for the packaging of orange juices, while the homogenization was done by the direct addition of the antioxidant to a mixing tank (60 L) for 5 min. Heat treatment of juice samples was performed using a pilot tubular pasteurization system (SPX Flow Technology, MX-0309124, 2010). Subsequently, the samples of the orange juice elaborated with 0.10 g/L of


Table 9.4  Color Parameters of an Orange Juice Supplemented With Antioxidant at Lab Level Color Parameter Treatment






Fresh juice Treated juicea (60°C × 30 min)

50.52 ± 0.02 50.96 ± 0.01

0.49 ± 0.08 0.88 ± 0.00

34.38 ± 0.00 34.38 ± 0.08

0.014 ± 0.00 0.025 ± 0.00

34.38 ± 0.001 35.14. ± 0.08


Supplemented with antioxidant (0.5 g/L).

a­ ntioxidant were treated with a processing temperature of 90°C and 30 s of residence time. After pasteurization at pilot plant level of an orange juice (formulation not specified), samples were stored at refrigeration for 24 h until further analysis (unpublished data). During the microbial count analysis performed, the samples obtained nondetectable counts for Escherichia coli, coliforms, Salmonella, and Staphylococcus aureus; in contrast, the sample without heat treatment (NTJ) presented significant microbial growths of 56 UFC/mL for deteriorating microorganisms (molds and yeasts) and 37 UFC/mL for aerobic mesophilic microorganisms where the formulated pasteurized juice (FPJ) did not. These results represent without doubts, the usefulness and the necessity to perform thermal treatments in orange juice to ensure the safety of the drinks and as a measure of quality assurance. Furthermore, the physicochemical analyzes (Table 9.5) showed that the AOA decreased with the thermal treatment (loose of 36.83%). This is of high importance due to the addition of antioxidant, result in high levels of AOA but this activity is lost with the thermal treatment, so another more gently treatment should be consider to avoid the loose of the levels of AOA. Related to heat treatments, Kathiravan et al. (2014) (Kathiravan et al., 2014) standardized a batch thermal pasteurization process with different total heating times (96°C/540 s, 96°C/720 s, and 96°C/900 s) and quantified their effect on antioxidants, pigments, and microbial inactivation of a ready-to-drink beet juice; they concluded that processing and storage had a decisive and significant impact on the degradation in betalain content, color, and AOA. On the other hand, the use of a protected antioxidant added to pasteurized infusions led to a caloric increase about a 6% due to the use of an encapsulating oligosaccharide. However, this change was perceived by a taster in these type of beverages (data not shown)


Table 9.5  Physicochemical Analyses of an Orange Juice Supplemented With Antioxidant at Pilot Plant Level Treatment Parameter




Antioxidant activity pH Acidity Moisture Total soluble solids L* a* b*

(%) (dimensionless) (g/L) (%) (°Brix) (dimensionless) (dimensionless) (dimensionless)

90.10 ± 6.89 3.48 ± 0.01 1.79 ± 0.01 91.26 ± 0.24 8.90 ± 0.01 54.34 ± 0.81 −3.47 ± 0.47 34.05 ± 0.62

53.27 ± 0.11 3.43 ± 0.01 1.98 ± 0.09 90.71 ± 0.54 8.65 ± 0.21 48.09 ± 0.37 −2.34 ± 0.02 33.10 ± 0.24

FPJ, formulated pasteurized juice; NTJ, nontreated juice.

and has to be taken into account due to the consumers’ demands and to the current requirements low calorie and healthy products. These samples did not present changes the response variables of pH, acidity, total soluble solids, water activity, or viscosity. However, the effect of pasteurizing represents high changes with respect to control in AOA. The most relevant physicochemical response variables were due to AOA and colorimetric response properties. The colorimetric measurements show a decreased in the luminosity color parameter (L*) of 6.25 units, while the color changes in a* and b* parameters were not pronounced (about one unit each). This is of vital importance due to the color is the most important attribute of visual quality for consumers, so a darkening could reduce sales because customers do not perceive a fresh product when compared to a recently prepared one. However, alternative and more innovative processes are nowadays available for beverage industry and as a result, cold packaging has become a very popular technology, and other technologies like high-pressure processing (HPP) (Chen et  al., 2015; Xu et  al., 2015; Swami Hulle and Srinivasa Rao, 2016), pulsed electric fields (PEFs) (Zulueta et al., 2010; Agcam et al., 2016; Carbonell-Capella et al., 2017), or UHT (Alqahtani et al., 2014) that have showed minimal affectation in beverages quality (see Section 9.5).


9.4  Sensorial and Analytical Analysis Nowadays, consumers are making stronger demands on the quality of processed foods, including health benefits, looking for flavors, aromas, and colors more related to the natural products. As a consequence, the food industry is taking special attention to avoid sensorial and organoleptic changes in its processed products including taste and aromatic profiles of different kinds of fruits (Jambrak et al., 2017). In this sense, novel preservation technologies to avoid loss of freshness as PEFs or high hydrostatic and dynamic pressures have been proposed. Nevertheless, there are still some factors that these nonthermal technologies need to overcome (CerdánCalero et al., 2013). For this reason, among the different existing processing technologies, heat pasteurization is the most common technique to extend the shelf life of natural products, due to the better inactivation of spoilage microorganisms and thermally resistant endogenous enzymes (Wibowo et al., 2015a). On the other hand, food industry is also using strategies that include sexual hybridization to develop new products, as an example, Barboni et al. (2009) reported the hybrid between mandarin and clementine trees, to increase the resistance against diseases and climatic conditions, obtaining fruits with flavors in adequacy with the consumer’s taste. They also produced fresh juices that preserve the organoleptic properties of the clementine fruit (Barboni et al., 2009). It has been well reported that the flavor, together with the appearance and texture, is crucial attributes for the acceptance and consumption of a particular food (Mastello et al., 2015). In addition, some authors have also reported that the general composition and antioxidant properties of foodstuff were major factors in determining consumer acceptance and preference. Fresh juices comprise a complex mixture of phytochemicals, organic acids, and phenolic compounds, which represents among the major compounds of citrus fruits (Kelebek et al., 2009); some others are volatile compounds that give them its characteristic aroma and taste. A part of these compounds can also contribute with biological activities, but only a portion of the volatile compounds are odor active, nevertheless they all collectively contribute to the overall perceived odor (Breksa et al., 2009; Mastello et al., 2015). The aroma of freshly squeezed orange juice has been attributed to aldehydes, mainly acetaldehyde, hexanal, octanal, and decanal, some esters especially ethyl butanoate, terpenes such as myrcene, alfa-pinene, limonene, some alcohols, and ketones (Mastello et  al., 2015). In the recent years, more attention had been paid on phenolic compounds including flavanones of citrus fruits, due to its antioxidant activities. In orange juices, the most common polyphenols identified are phenolic acids as hydroxycinnamic acid and its derivatives (ferulic, p-cumaric, sinapic, caffeic, and chlorogenic acids). Flavanones mainly occur as glycosides, hesperidin, and narirutin, known as the main flavanones, but it is possible to find also didymin, neohesperidin, and naringin in orange juices (Kelebek et al., 2009).


One of the most difficult challenges specifically in orange juice industry is to retain the flavor close to that of freshly squeezed juice to guarantee consumers a consistent natural flavor (Mastello et al., 2015). The quality of the product during storage is also a big challenge, due to orange juice changes in sensorial and nutritional qualities through time, determining the best date listed on the product (Wibowo et al., 2015a). In the juice industry, the processed products require simple and rapid procedures (Barboni et al., 2009), the most common analysis to evaluate organoleptic parameters is the sensory analysis, which is performed by trained panelists (Jambrak et al., 2017). The juice industry has also developed specific analytical techniques to allow the identification and determination of abundant flavors and off-flavors, which affect directly the sensorial quality of products (Barboni et al., 2009). Furthermore, several techniques to identify polyphenols, flavonoids, and terpenoids, among other phytochemical compounds related to the sensorial attributes have been also described.

9.4.1  Sensory Analysis of Citric Fruit Juices Flavor is describe as one of the most multisensory aspects of our everyday experience, its perception brings together different stimuli: aroma, taste, texture, and some esthetic sensations, but also sound and visual cues (Charles et al., 2017). Fruit aroma is a complex combination of numerous volatile compounds that contribute to the overall sensory quality of fruit specific to species and varieties (Charles et al., 2017). The volatile profile is generally chemically/instrumentally detected, but does not necessarily reflect the aroma profile of the sample, making important the use of supplementary methods to identify the odor-active compounds responsible for the overall aroma (Mastello et al., 2015). In this sense, different methodologies for sensory analysis have been reported. Charles et  al. (2017) studied the relations between aroma, taste, and texture in a complex matrix of juices, using the dynamic sensory method (TDS) as a part of an original approach in addition to sensory and instrumental static methods. Other comparisons tests have been applied in independent sessions, to evaluate the sensorial acceptability of samples according to the unstructured scales (CerdánCalero et al., 2013). Furthermore, free-choice profile (FCP), a method developed in 1980 that can be carried out by untrained panels that use just a specific scale of an attribute with data generated by sophisticated statistical methodologies, has been used for sensorial analysis in processed and storage juices (Pérez Aparicio et al., 2007). Pérez et  al. (2007) reported the use of this method (FCP) to generate the descriptors from different orange juice types using a robust principal components analysis (PCA). The authors evaluated (1) fresh orange juices with different storage days (not specified), (2) pasteurized (and


concentrated) samples, and (3) orange nectar drinks from different brands. The results indicated that this methodology represents a useful tool in marketing decisions and in quality-product program development. Even though sensorial analysis using trained or untrained panelist is the most common methodology used in the juice industry, sometimes it takes large periods of time when trained panelists are required or when multiple samples must be evaluated, in this sense several methods have been developed to facilitate the analysis.

9.4.2  Analytical Methods of Citric Fruit Juices The threshold of perception of the fragrant compounds that contribute to the principal sensory characteristics of the product can change depending on whether the volatile compound is present alone or in combination with others (Barboni et al., 2009). Different extraction processes are the main scientific focus not only to recover phytochemical compounds for better use in food industry but also to solve an environmental problem. Organic acids, sugar, and phenolic compounds are also presents in citrus fruits and its nature and concentration largely affect taste characteristic and organoleptic quality (Kelebek et  al., 2009). As already mentioned the considerable interest and attention to the polyphenol compounds due to their bioactive functions has derived in many efforts to provide a highly sensitive and selective analytical method to its determination (Ignat et al., 2011). For this reason, technical expertise and sophisticated equipment are important factors to evaluate the quality standards of chemical, physical, and sensorial parameters of juice product using analytical techniques (Jambrak et al., 2017). Two principal factors are essential to obtain and suitable analytical analysis: (A) sample preparations that involves extraction on the desirable compounds and (B) adequate selection of identification and/or quantification method. For sample preparation in volatile compound extraction, several techniques have been proposed. Cerdán-Calero et  al. (2013) have reported extraction with fiber coated with polydimethylsiloxane (PDMS) as an efficient sample preparation. Preconcentration of volatile compounds by solid-phase micro-extraction (SPME) and solid- phase micro-extraction in headspace mode (HS-SPME) has been also well appreciated for its characteristics. Specific HS-SPME is nowadays the most suitable techniques of sample concentration, due to its simplicity, rapidity, reliability, and the absence of solvent that does not induce modifications of volatile compounds due to temperature or solvent effect (Barboni et al., 2009; Jambrak et al., 2017). For polyphenol extractions, several techniques have been reported, generally these compounds have been extracted by gridding, drying or ­lyophilizing fruits, or by soaking fresh material with subsequent


solvent extraction (Ignat et al., 2011). Nevertheless, these methodologies (solid-phase (SPE), liquid-liquid, and solid-liquid extraction) imply the co-extraction of nonphenolic compounds and subsequent purification process is required. Environmentally beneficial alternatives have also been proposed as supercritical fluid, microwave, and enzymatic extractions (Ignat et al., 2011). In the identification and quantification of volatile compounds, several techniques have been developed using gas chromatography (GC) and GC coupled with a mass spectrometer (GC-MS), this methods have been used for the analysis of odors such as tomato, apricot, pear, peach, strawberry, apple, oranges, and mandarin (Barboni et al., 2009; Jambrak et al., 2017). For identification and quantification of polyphenols and terpenoids in citrus fruit juices, different types of chromatography have been used: (1) thin-layer chromatography (TLC), (2) high-pressure liquid chromatography (HPLC), (3) capillary electrophoresis (CE), (4) liquid chromatography coupled to mass spectrometry (LC-MS), and (5) LC-electrospray ionization mass spectrometry (LC-ECI-MS), methods that have all been described in the literature (Breksa et al., 2009). The HPLC chromatography with a diode array detector (DAD) has been the most used system to polyphenols quantification. Mobil phases as water, acetonitrile and formic acid, as well as different concentrations of gradients to the separation and identification of the majority of the compounds in citrus juice samples have been also evaluated, the ­ultra-violet-visible spectra reported in those studies are in the scanning from 200 to 600 nm (Kelebek et al., 2009). Moreover, a satisfactory ultra performance liquid chromatography (UPLC)-DAD methodology has been developed to the quantification of the main polyphenols present in citrus fruits (CIATEJ’s Chromatography Laboratory). In Fig.  9.6, a chromatogram of a polyphenol mixture was obtained by the use of a Waters UPLC Acquity H Class chromatographer (Milford, MA, United States) equipped with a quaternary pump (UPQSM), autosampler injector (UPPDALTC), and

Fig. 9.6  UPLC chromatogram of polyphenol standards obtained by a DAD detector.


DAD eλ photodiode array detector (UPPDALTC). Empower 3 software (Waters, Milford, MA, United States, 2010) was used for data acquisition and processing. Chromatographic separation was carried out using a Waters Acquity UPLC BEH C18 column, 1.7 μm, 100 × 2.1 mm I. D. (Milford, MA, United States) at 25°C temperature, the flow rate was kept at 0.2 mL min−1 and the injection volume was programed at 2 μL. The developed methodology allowed the quantification of the mixture of 13 polyphenols present in citrus fruits. A clear definition of the peaks for each compound was obtained. Chlorogenic and caffeic acid eluted during the first 9 min, then rufin, eriocitrin, ellagic acid, p-­coumaric acid, and sinapic acid eluted between 9 and 10 min, from 10.3 to 11.5 min the presence of diosmin, naringin, hesperidin, and neohesperidin was observed. Finally, at 13.4 and 16.15 min, morin and quercetin were observed, respectively.

9.4.3  Principal Results of Analytical Analysis of Aromatic Profiles and Polyphenol Contents in Citrus Beverages Among the results reported in the literature, Barboni et  al. (2009) performed the analysis by HS-SPME-GC and/or GC-MS of the volatile compositions of 67 juices obtained from mandarin, clementine, and hybrids fresh fruits to allow the identification of 44 compounds. The clementine juice presented predominance of limonene (90%) and low presence of γ-terpinene (1.2%), while mandarin juice exhibits high amount of limonene (66.3%) and γ-terpinene (21%) (Barboni et  al., 2009). Cerdán-Calero et al. (2013) reported the high-pressure homogenization (HPH) at 150 MPa of centrifuged partially depulped juices as an alternative to thermal pasteurization process; this methodology resulted in a product with fresh taste, color, cloudiness, and aroma composition close to those of the original fresh juice. A GC-MS analysis with automated data processing performed by the automated mass spectral deconvolution and identification system (AMDIS) was proposed by the authors as a new methodology to facilitate data analysis. A total of 88 volatile compounds and the evolution of 32 aroma descriptors were able to analyzed, the comparison of the process and aroma profiles were also determined. These results allowed identifying that the homogenized juice was initially closer to the fresh one, and presented lower concentrations in off-flavor compounds during storage; information generated with used just sensorial panelists, was difficult to achieve. Kelebek et  al. (2009) reported the use of HPLC methods to identify and quantify organic acids, sugars, and phenolic compounds in orange juice obtained from the cv. Kozan of Turkey. Three organic acids (citric, malic, and AAs) and three sugars (sucrose, glucose, and fructose) were determined. Moreover, a total of 13 phenolic compounds were identified, where the flavanones hesperidin and narirutin were the most abundant.


The determination of the most common polyphenols present in orange juice was performed (CIATEJ’s Chromatography Laboratory) using four different commercial samples (three of them from Citrus sinensis and one from Citrus aurantium) and the UPLC-DAD methodology described in the previous section. The results (Table 9.6) indicated the presence of hesperidin in all C. sinensis samples as the major polyphenol at concentrations from 23 ± 0.7 to 423 ± 1.1 μg/mL, they also presented sinapic acid and just one of them presented neohesperidin. The commercial sample of bitter orange juice presented neohesperidin as the major polyphenol (1221.8 ± 2.1 μg/mL), followed by naringin, p-­coumaric acid, hesperidin, chlorogenic acid, and caffeic acid. This represents a high level of total antioxidants (1630 ± 1.98 μg/mL) maybe due to a high proportion of antioxidants in the formula (Table 9.1) as citric or AAs.

9.4.4  Emerging Technologies in Sensorial and Analytical Analysis Due to the substantial amount of resources, time, and costs that sensorial analysis requires (Jambrak et  al., 2017), new methodologies including the use of electronic tongues of several types (potentiometric, ­voltammetric, and impedance) may represent low-cost and reliable alternatives. Some systems have been used to test several juices with a combination of voltammetric electronic tongue and a gas sensor array (Jambrak et al., 2017). Equipment as electronic nose and tongue has been successfully applied for semiquantitative discrimination of drinks (Cerdán-Calero et al., 2013; Jambrak et al., 2017). Mastello et al. (2015) reported the use of GC-olfactometry (GC-O) employing heart-cut multidimensional GC techniques with olfactometry (O) and mass spectrometry (H/C MDGC-O/MS) and comprehensive two-dimensional (2D) GC-accurate ass time-offlight MS (GCxGC-accTOFMS) to analyze odor-active compounds, they founded that the multiassessment instrument approach employed provided an effective insight into the processed orange juice aroma.

9.5  Effect of Storage Conditions on Product Shelf Life 9.5.1  Shelf Life Nowadays, the nonalcoholic beverage industry centers in producing safe, high-quality, and healthy products for their customers. To achieve this goal, they add active compounds like vitamins or antioxidants to offer functional beverages and also, this industry considers shelf life an important issue to offer to customers as part of its quality. Shelf life is defined as the time during which an edible product r­emains safe to consume, and retains its chemical, physical,

Table 9.6  Polyphenol Analysis in Commercial Orange Juices Samples Compounds (μg/mL) Hesperidin



Total Polyphenols (μg/mL)

Nd Nd 331.3 ± 0.5

422.9 ± 1.1 87.1 ± 0.3 23.1 ± 0.7

1.2 ± 0.01 Nd 1221.8 ± 2.1

Nd Nd 1.1 ± 0.01

430.0 ± 0.75 93.8 ± 0.58 1630.9 ± 1.98


343.4 ± 0.2



350.2 ± 0.06


Chlorogenic Acid

Caffeic Acid

p-Coumaric Acid

Sinapic Acid



Nd Nd 20.1 ± 0.1

Nd Nd 8.3 ± 0.1

Nd Nd 25.4 ± 0.1

5.9 ± 0.3 6.8 ± 0.3 Nd





6.8 ± 0.3

BONO, Bitter orange juice, Noble; NP, nondetectable; ORCP, Orange juice, Cosecha Pura; ORDV, Orange juice, Del Valle; ORJU, Orange juice, Jumex.


­ icrobiological, nutritional, and sensorial characteristics when stored m under recommended conditions, complying with label declaration (Institute of Food Science and Technology (UK), 1993). Therefore, shelf life is a function of time and environmental factors during storage (Giménez et  al., 2012); in nonalcoholic beverages, these environmental factors are temperature and light, although the process of production has an important role in the shelf life’s duration. Citric juices characteristics like color, vitamin C concentration, flavonoids, the presence of microorganisms, and the activity of food additives or nutraceutical are affected by time, temperature, and light. A crucial quality loss parameter on orange juices during shelf life is the change in color, this is the first visible sign which can negatively influence the consumer’s acceptance that decreases its commercial value (Wibowo et  al., 2015b). Color is an important characteristic in citrus juices, plays a role as a quality indicator and in consumers’ preferences, because of this, its stability has been studied in different citrus fruits. For instance, pasteurized orange juices were tested overtime at different storage temperature conditions for evaluating nonenzymatic browning and the carotenoid chemical changes. The nonenzymatic browning study, reports that orange juice components like sugars, AA, oxygen presence, acidity change, furfural, and HMF are the main factors responsible in color changes in pasteurized orange juice, since they show significant changes in concentration as a function of time and temperature (Wibowo et  al., 2015a). In the case of carotenoid chemical changes, results from this study indicated that carotenoid profile showed important changes and some of them appeared to have different susceptibilities to storage conditions, and the isomerization of these pigments was more important than their own oxidation reactions. In addition, orange juice contains an appreciable amount of AA, so the authors suggest that AA degradation may contribute to browning (Wibowo et al., 2015b). In the same context, color intensity during shelf life of bloody orange juice degassed and fortified with AA was evaluated at different storage time and different storage temperature. The results of this study showed a color intensity degradation as a consequence of the storage time and temperature; however, the degassing retained more color intensity during storage. However, AA fortification at tested concentrations has no effect on the rate constants of intensity color loss (Remini et al., 2015). Moreover, it is important to highlight in these reports, the relation between both color and AA stability. Burdurlu et al. (2006) evaluated the AA degradation in citrus juices concentrates of orange, lemon, grapefruit, and tangerine during different storage times and temperatures. They report that AA in citrus juice concentrates decreased with increasing storage temperatures and times, and decomposes easily in acid solutions, being the lemon juice concentrates (pH 1.82) the ones that showed the highest AA destruction.


Commonly during processing, the commercial citrus juice is pasteurized to (1) avoid the presence of microorganisms, mainly the foodborne pathogens, and (2) to reduce pectin methylesterase activity, which affects the cloud stability and viscosity of citrus juice by de-esterification of the methoxylated pectin (Navarro et al., 2014; Aghajanzadeh et al., 2016). Nevertheless, the heat affects AOA, color, and nutritional components in citrus juices decreasing their quality and healthiness (La Cava and Sgroppo, 2015; Aghajanzadeh et  al., 2016). For instance, after the thermic treatment in a nutraceutical orange juice, formulated for a Mexican company at pilot plant level (supplemented antioxidant: 0.10 mg L−1; pasteurization parameters: 90°C/30 s product, previous section), the total color change (ΔE*) and the AOA were evaluated as their quality parameters during 5 weeks of shelf life at different storage temperatures. Both parameters presented a zero-order model (Fig. 9.7), showing a less complex kinetic of 25





2°C 30°C


45°C 0








Time (weeks)


Antioxidante activity (%)

100 80 60 40

2°C 30°C


45°C 0




2 Time (weeks)



Fig. 9.7  Effect of the time and temperature storage in pasteurized nutraceutical orange juice. Measured data (symbols), and zero-order kinetic model (lines) of: (A) total color intensity (ΔE*) and (B) antioxidant activity (%).


­ egradation. Furthermore, the shelf life calculated for the orange juice d was determined of 2.4 and 18.1 weeks for ΔE* and AOA, respectively, ­taking a reduction of 50% as the critical value for AOA, and for ΔE* a value of 2.8 units as a value proposed recently because the change of color is visually distinguised by people in orange juice (Wibowo et al., 2015b). These values lead to acceptable shelf life for supplemented orange juice. However, further studies should be done in thermal pasteurization to increase the shelf life with high storage times and higher values of stable AOA without degradation. On the other hand, the calculated activation energy (Ea) was 64.68 ± 0.70 kJ mol−1 and 74.12 kJ mol−1 for ΔE* and AOA, respectively (Table 9.7). The Ea is related with the velocity of degradation, the higher activation energy from an orange juice implies that a small temperature change is needed to degrade color or AOA more rapidly (Burdurlu et al., 2006; Remini et  al., 2015). Wibowo et  al. (2015b) report the same kinetic model for ΔE* with a value of Ea of 69.83 ± 1.76 kJ mol−1 in orange juice pasteurized in similar conditions, while Remini et al. (2015) measured the anthocyanins content in the juices and reported a second-order model for color intensity loss, with a Ea of 58 ± 28 and 99 kJ mol−1 for moro

Table 9.7  Activation Energy (Ea) of Total Color Intensity and Antioxidant Activity (%) of Pasteurized Orange Juices. Product Pasteurized nutraceutical orange juice Pasteurized orange juicea Pasteurized moro blood orange juice fortified with ascorbic acid (100 mg L−1)b Pasteurized moro blood orange juice fortified with ascorbic acid and deaeratedb a

Pasteurization Condition


Ea (KJ·mol−1)

Shelf-Life at 25°C (Weeks)

90°C, 30 s 80°C, 2 min

ΔE* Antioxidant activity ΔE* ΔE*

64.68 ± 0.70 74.12 ± 1.24 69.83 ± 1.76 58 ± 28

2.4 18.1 nr nr

80°C, 2 min


99 ± nd


90°C, 30 s

Wibowo et al. (2015b). Remini et al. (2015). nd, not determined; nr, not reported.



blood orange juice fortified with AA (100 mg L−1) and for fortified and deaerated juice, respectively; however, they did not measured ΔE*.

9.5.2  Alternative Techniques The damage caused to citrus juices by heat has attracted research into alternative techniques to reach this objective. Nonthermal technologies for food processing have been receiving great attention due to the ability to improve the quality and safety of food. UV-C light and HPP are some examples. For instance, Torres et al. (2011) applied HPP to quantify the shelf life of bloody oranges juices after the HPP treatmeant at 400, 500, and 600 MPa for 15 min. The AA and anthocyanins stability was determined as quality parameters, during 10 days of storage at 4 and 20°C. This report indicates high levels of anthocyanins retention quantified as cyanidin-3-glucoside with a shelf life at 20°C of 84.8 days due to the AA during HPP at 600 MPa for 15 min, while in a control sample was 20.1 days. However, no significant differences were observed between 600 MPa/15 min processed and control samples during storage at 4°C (Torres et  al., 2011). The authors also pressurized to 402 ± 1.9 MPa for 3 min a grapefruit juice and stored it for 21 days at 4°C evaluating the most critical quality parameters as color, organic acids (AA and citric acid), flavonoids, coumarins, carotenoids, and the presence of microorganisms. The results of this study show that HPP treatment maintained the levels of phytochemicals as flavonoids, limonoids, coumarins, and citric acid, all of them with no significant variation comparable to the control sample and pasteurization treatment during stored at 4°C. However, grapefruit juice treated with HPP had better stability for AA and the color parameters L* and a*, than the pasteurization treatment during the storage. Microorganism presence during storage was below of detectable limits in both treatments, HPP and pasteurization (Uckoo et al., 2013). On the other hand, an UV-C light treatment was applied to grapefruit juice at different doses (0.0–3.94 J/cm2). The effect in the main bioactive compounds and their stability were evaluated throughout 30 and 16 days of storage at 4 and 10°C, respectively. After UV-C treatment, AA and antioxidant capacity decreased significantly, which was related to the applied dose. However, no changes were observed in other organic acids, in phytochemicals as flavonoids and total phenols, or in physicochemical parameters like pH, °Brix, color, and titratable acidity after UV-C treatment. During the shelf life at both temperatures, a decrease in the neohesperidin and total phenols levels was detected, whereas the others parameters analyzed did not show any changes. In addition, at both temperatures of storage, the samples treated with UV-C showed a decrease in the growth rate constant for total aerobic microorganisms and yeasts and molds compared with untreated grapefruit juice (La Cava and Sgroppo, 2015).


In addition, the application of thermosonication (TS) and PES in citrus juices has been reported (Walkling-Ribeiro et  al., 2009); however, after 168 storage days, color shelf life and microbial counting were better with the traditional pasteurization method.

9.6  Antioxidants and Improvement of Antioxidant Beverage Power: Alternative Sources and Costs 9.6.1  Alternative Sources Citrus juices are one of the main sources of vitamin C and AA in the human diet, its importance is noticeable in AOA but it is an unstable compound and under less desirable conditions it decomposes easily, therefore, its concentration and stability must be taken into consideration to offer a safe and healthy product; hence, it is important to continue the research in innovative methods to allow a stable AA during storage and a prolongated shelf life, since its presence affects the rest of components in citrus juices. Besides the addition of vitamin C, as pointed out, other antioxidants compounds like polyphenols, flavonoids, or carotenoids are under study to be used in food industry in order to improve quality and cover the market demand. These alternative compounds can be obtained from agro-industrial wastes or under exploited fruit or vegetables. Strawberries, blackberries, mango’s peel, pomegranate, bananas, and other endemic fruits have been used for this purpose. Pomegranate (Punica granatum) has important concentrations of phenolic compounds, flavonoids, anthocyanins, tannins, AAs, and gallic acid. Pomegranate peel dried extract was added in different concentrations to tomato juice and orange-strawberries juice in order to improve the antioxidant content of these beverages. Results showed that the AOA was increased in both juices compared to controls as much as concentration of peel extract was added. The lower concentration tested (0.5%) was better accepted according to the sensory analysis, though (Mastrodi Salgado et al., 2012). Banana (Musa acuminata Colla AAA) peel is the main biowaste of the banana processing, besides being a source to obtain different compounds like protein, cellulose, pectin, and enzymes, it is rich in phenolic compounds, anthocyanins, carotenoids, sterols, triterpenes, and catecholamines. Banana peel extract (BPE) was used to increase AOA in orange juice, fresh squeezed and from concentrate. Results showed that the scavenge free radicals of juices were increased when BPE was added in all concentrations. However, sensorial tests indicated that concentrations over 10 mg BPE/mL were detected in mouth and an unpleasant odor (Ortiz et al., 2017).


Jamun (Syzygium cumini) or Indian blackberry is an indigenous fruit with short shelf life, astringent flavor, and rich in phytonutrients (anthocyanins, tannins, alkaloids, terpenoids, and minerals). The jamun processed as a powder was added to pear juice in order to increase its antioxidant content and as expected, results were positive. A 4% supplementation was the best concentration accepted after sensorial tests, also increasing 18.13% the AOA in pear juice (Kapoor and Ranote, 2016). Besides, Kapoor and Ranote (2016) used freeze drying and hot air drying, concluding the former was the best method to process jamun extract due to bioactive compounds were less affected, despite drying process time was 60 h. Other alternative sources of bio-compounds already been studied to be added in beverages are: (1) extracts from citrus peel or seed (Rafiq et al., 2016); (2) by-products of cashew Anacardium occidentale (Ajileye et  al., 2015; Gastélum-Martínez et  al., 2016; Das and Arora, 2017); (3) mango peel (Dorta et al., 2012; Torres-León et al., 2017); (4) grape fruit peel (Farhadi et al., 2016; Pantelić et al., 2016); and (5) pink guava (Rojas-Garbanzo et al., 2016), just to mention some. A different alternative in supplementation was developed by CocaCola North America (Apopka, FL), in which microencapsulated algal oil high in docosahexaenoic acid (DHA) was added to orange juice manufactured in order to evaluate the effects in plasma phospholipid DHA content in children from 4- to 6-year-old and 7- to 12-year-old. Results showed that supplementation was effective in increasing DHA contents and as an effective mean of enhancing DHA intake in children (Hawthorne et al., 2009). As we could see, besides AA there are other sources of antioxidants to be added to beverages. However, further studies are needed to achieve optimal extraction of these biocomponents and to ensure their activity in vivo after consumption.

9.6.2  Cost/Benefit of Adding Antioxidants Other important aspects to consider in the addition of antioxidants to juices or beverages are the product final cost and the consumer’s willingness for paying the benefits claimed in the product. Product final cost might be highly affected by the process complexity to obtain the antioxidants compounds to be added in the beverage. An ideal recovery process should have: – As less extraction process steps as possible – Short processing time – Availability for scaling-up – As less purification process steps as possible – High yields – High stability (tolerance to heat, light, pH…) – Demonstrated bioactivity – Eco friendly process


All these conditions in order to avoid considerable extra costs increment in final price. Finally, a profound understanding of the bio/pharmacokinetics will help to unravel the compounds that can ultimately lead to true, established nutraceuticals, while simultaneously allowing for the establishment of relevant legislation and efficient regulation. From the industry’s perspective, such research will give credit to the health claims and will identify which compounds are, in fact, nutraceuticals worthy for commercialization, and, maybe, lead to “healthier” profits (da Costa, 2017).

9.7 Conclusions As we reviewed through this chapter, antioxidants are important when elaborating an orange/citric juice due to the healthy profile that the beverage acquired, the microbial security of the beverage, and the healthy appearance between the consumers. Ingredients and labeling has a critical importance in orange juice, the inclusion of stabilizers for providing them of consistency is relevant; as showed the greater viscosity presented was due to an bitter orange juice; however, consistently all juices presented a Newtonian behavior. The addition or supplementation of beverages needs to consider several steps like the selection of ingredients (antioxidant compounds, mainly), formulation, and characterization. In the formulation step, the most used antioxidants are the citric acid, followed of malic, tartaric, or AA in juices. Nowadays, some citric juices and concentrates in the market have troubles to reach standard values without high variability. Then, without doubts, the need of a standardized processing should be a mandatory task in every beverage company. The antioxidant addition has to be considered: some of the commented antioxidants as polyphenols are expensive and some are degraded within the thermal treatment. Different sources of antioxidants are now available as citric, banana, pomegranate, and grapefruit peels wastes. When a beverage is added with compounds extracted from alternative sources, their stability must be ensure, especially if a thermal treatment is included during the processing in order to guarantee a good final antioxidant capacity. Extraction of hesperidin from citrus waste is an option to get polyphenols. The encapsulation with CDs was effective to preserve AOA of hesperidin in supplemented citric beverages. Sensorial analysis is a useful tool to identify specific flavors and odors in fruit juices that combined with analytical methods can provide useful information for the acceptation of consumers.


An acceptable shelf life for thermal treated and supplemented with antioxidants orange juices was calculated (18 weeks) with the AOA; however, a short shelf life due to color parameter was presented in this juice. Novel technologies discussed are difficult to scale-up and have an economic impact in final product costs, as a consequence pasteurization is still a useful thermal-preservative methodology, even when an antioxidant is added.

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Azime Özkan-Karabacak, Bige İncedayı, Ömer Utku Çopur Uludag University, Faculty of Agriculture, Department of Food Engineering, Bursa, Turkey

10.1 Introduction Thermal processes could be mainly used in preservation of beverages to enable the microbiological and enzymatic inactivation. However, high process temperatures gain negative effects on color, nutritional value, taste, and sensorial properties of beverages. Nowadays consumers pay more attention to minimally processed and fresh food products without changed organoleptic and nutritional attributes besides purified from additives, preservatives, and environmental impurities (Thakur and Nelson, 1998; Raso and Barbosa-Canovas, 2003; Yamamoto, 2017). Because of these reasons many food producers and consumers have been interested in nonthermal preservation technologies (Tao et  al., 2015; Tokuşoğlu, 2016). The fundamental aims of these technologies are to decrease the processing times and to improve the process conditions. They also have an excellent inactivation effect on microorganisms and enzymes when used with combined thermal processes (Norton and Sun, 2008). The current studies are based on novel nonthermal techniques, particularly the treatment of high hydrostatic pressure (HHP) on shelf life prolonging of foods. Moreover, HHP offers advantages in terms of processing time, quality, and energy and minimizes the chemical and physical hazards in foods (Oey et al., 2008; Patterson et al., 2008; Barrett and Lloyd, 2012). HHP is also gained an acceptance as a safety step in beverage processing and have a high consumer demand for safe and nutritious fruit and vegetable juices, dairy beverages, etc. (Koutchma, 2014). Nowadays, application of HHP and its effects on beverage nutrients have been greatly studied. This chapter highlights the challenges and opportunities relevant to using HHP technology in beverages and makes proposals to producers and consumers on application of this method. Preservatives for the Beverage Industry. © 2019 Elsevier Inc. All rights reserved.


310  Chapter 10  Preservation of Beverage Nutrients by High Hydrostatic Pressure

10.2  The System of HHP The abbreviations of high pressure technique are altered in the literature as high hydrostatic pressure processing (HHP, HPP, HP) or high pressure homogenization (HPH). The main difference among the names is associated with application methods (Icier, 2012). After Hite (1899) had been suggested to apply HHP on raw milk to extend its shelf life at the end of 19th century, the researches and developments on HHP food processing have become popular and spread out worldwide (Farkas and Hoover, 2000; Yamamoto, 2017). Thereafter in the 1990s, HHP applications in food industry progress rapidly due to the benefits on inactivation of enzymes and microorganisms by preserving nutritional value of foods more than conventional treatments (Tokuşoğlu and Doona, 2011). Exposure time, temperature and pressure, type of the packaging, and product parameters are the main properties affecting HHP on food. Instantaneous and isostatic pressure are nearly transmitted to the product without depending on the shape, size, and food composition (Koutchma, 2014). Moreover, extremely homogenous products can be improved with HHP (Deliza et al., 2005). The applied pressure is changed between 2000 bar (200 MPa) and 9000 bar (900 MPa) related with the capabilities of the HHP unit with or without extra external heat (Smelt et al., 2001; Koutchma, 2014; Koutchma et al., 2016). However, the accepted pressure to inactivate vegetative microorganisms is ranged from 4000 to 6000 bar (Smelt et  al., 2001). Table  10.1 shows frequently used pressure units in HHP application and their conversion. A uniform distribution of the pressure in all directions is the positive effect of HHP on beverages (Oey et al., 2008). Therefore, HHP-treated foods or beverages retain their original shape (Tokuşoğlu and Doona, 2011). In a typical HHP system, the water is generally used as the incompressible transmission fluid due to the slight decrease under pressure compared to gases. Food-grade solutions such as ethanol, sodium benzoate, castor oil, silicone oil, and glycol can be also used as the transmitting medium (Tao et al., 2015). The system comprises a closed HHP vessel, a pressure generation system, a temperature control apparatus, and a product usage system (Rastogi et al., 2007; Yaldagard et al., 2008). Before processing, packed foods (commonly in resilient plastic bag material or plastic bottles) are filled into trays which are conveyed to the water loaded vessel. After the vessel is covered and sealed, pressure at required level is applied. Samples are exposed to pressure at desired times (generally 1–5 min) (Icier, 2012). Pressure can be formed by different methods such as direct compression (used in lab scale systems), indirect compression (used in

Chapter 10  Preservation of Beverage Nutrients by High Hydrostatic Pressure   311

Table 10.1  Frequently Used Pressure Units and Their Conversion 1 Pa = 1 N/ m2 1 bar 1 psi 1 atm

Pascal (Pa)

Bars (bar)

Pounds per square inch (psi)

Atmosphere (atm)





100,000 6894.76 101,325

1 0.0689 1.01325

14.504 1 14.6959

0.9869 0.0680 1

From Koutchma, T., 2014. Adapting High Hydrostatic Pressure (HPP) for Food Processing Operations. Academic Press, with permission.

commercial applications), or heating pressure medium (applied for the treatments where elevated pressure in combination with high temperatures are necessary). In direct method (Fig.  10.1), a piston is needed and squeezing process is particularly fast. The sealing problem between the piston and interior surface of container concludes to use this method only in the laboratory-scale plants. The second method is more widespread which is occurred by using pressure amplifier to pump liquid from the pressure middle tank to the cell, until the r­equired pressure value is achieved (Fig.  10.2). The third one is ­especially useful when a combined effect of high temperature and pressure is required. This technique is not practically applied in food

Press frame Pressure vessel Piston Pressure medium

Bottom sealing

Fig. 10.1  Schematic diagram for the HHP using the direct method. From Bertucco, A., Spilimbergo, S., 2001. Treating microorganisms with high pressure, in: Bertucco, A., Vetter, G. (Eds.), High Pressure Process Technology: Fundamentals and Applications. Elsevier, Amsterdam, pp. 626–640, with permission.

312  Chapter 10  Preservation of Beverage Nutrients by High Hydrostatic Pressure

High pressure cell

Pressure medium tank

(max 1000 mPa)

P Water jacket


Fig. 10.2  Schematic diagram for the HHP using the indirect method. From Bertucco, A., Spilimbergo, S., 2001. Treating microorganisms with high pressure, in: Bertucco, A., Vetter, G. (Eds.), High Pressure Process Technology: Fundamentals and Applications. Elsevier, Amsterdam, pp. 626–640, with permission.

Pressure booster


industry (Thakur and Nelson, 1998; Bertucco and Spilimbergo, 2001). The effectiveness of HHP depends on the pressure level and duration rather than the size and amount of the product (Thakur and Nelson, 1998; Koutchma, 2014). The main pressurization systems are classified as batch and semi continuous. Batch systems can be used for both solid and liquid foods in industrial scale, whereas semi continuous systems recommended for the use in pumpable products such as beverages (Tao et al., 2015). Semicontinuous systems divided into two chambers by using free floating piston. While one chamber is used for the pumpable products, the other one is loaded with transmitting fluid. The pressure on the transmitting fluid is released after a convenient time. Then discharged liquid food from the vessel can be loaded aseptically into presterilized containers (Palou et al., 2002).

10.3  Used of HHP to Maintain Beverage Nutrients A wide range of beverages are consumed by people of all ages in daily diet especially to meet the nutrient requirement. There are many types of beverages, including fruit and vegetable juices, coffee, tea, milk, soda, alcohol (beer, wine), etc. (Wilson and Temple, 2016). The

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beverages such as fruit and vegetable juices are particularly preferred due to their bioactive compounds. Phenolic compounds, organic acids, and volatile compounds are secondary metabolites found in large amounts in plants. Antioxidant properties of bioactive compounds show anticarcinogenic effects besides the inhibition possibility of cardiovascular and cerebrovascular diseases (Nizamlıoğlu and Nas, 2010). With regard to nutritional perspective, HHP is a perfect technology for food processing which enables to retain healthy food components, color, and aroma profiles of beverages related with the low temperature treatments (Rastogi, 2013; Rawson et al., 2011). It is generally known that high levels of bioactive components were retained and limited detrimental effect of HHP on vitamins, pigments, and flavoring agents in the samples are realized due to its restricted influence on the covalent bond of low-molecular-mass components (Tauscher, 1998; Butz et al., 2003; Tokuşoğlu and Doona, 2011; Cao et  al., 2012). This effect is particularly important in beverages, which are abundant sources of antioxidants, pigments, and vitamins (Pandrangi et al., 2015). Moreover, the dissolution rates of the bioactives are increased by using high pressure extraction. Studies on preservation of phenolics by HHP confirmed that the concentration and extractability of these components were either decreased or not influenced after processing with HHP (Zhang et al., 2005; Corrales et al., 2008; Tokuşoğlu, 2016). The other advantage of HHP is to maintain vitamin stability of foods. Vitamin C is used as a quality index in fruits and vegetables due to its high sensitivity to heat than other bioactive components and therefore it acts as a current standard for other nutritional compounds (San Martin et al., 2002; Barba et al., 2012b). There are many studies on using HHP to maintain vitamin stability of beverages which are discussed in this chapter. It should be taken into consideration that enzymes are in charge of the stability of some bioactive components such as anthocyanins (Garcia-Palazon et  al., 2004). The response of enzymes exposed to HHP is changed based upon pressure, temperature, and time of treatment, the source of enzyme and structure of the substrates (San Martin et al., 2002). The impacts of HHP on protein denaturation change according to the kind of protein, treatment circumstances, and the pressure used. Protein denaturation is commonly reversible in 1000–3000 bar pressure range, but while the pressure is higher than 3000 bar, it becomes irreversible (Thakur and Nelson, 1998). The increase of pressure about 1000 bar causes an increment in the temperature of protein denaturation, while denaturation temperature generally decreases at higher pressures. Temperature and pressure combinations that caused ­protein

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Fig. 10.3  Pressure-temperature phase diagram of protein denaturation. From Messens, W., Van Camp J., Huyghebaert, A., 1997. The use of high pressure to modify functionality of food proteins. Trends Food Sci. Technol. 8, 107–112, with permission.

denaturation are limited by an ellipsoidal curve. Fig.  10.3 shows the ­pressure-temperature range in which protein preserves its native structure. The second area (II) represents under the maximal passing temperature, protein denaturation begins at lower temperatures using higher pressures. The third area (III) indicates that under the temperature relating to the maximal passing pressure, protein denaturation takes place at reduced pressures and temperatures (Messens et al., 1997; San Martin et al., 2002).

10.4  Effects of HHP on Different Beverage Nutrients 10.4.1 Fruit Juices Due to the minimizing the postharvest losses, there is a requirement to process fruit juices with the aim of shelf-life prolongation and retaining quality parameters of fresh fruit besides color, aroma, nutritional value, and structural properties. The major components of fruit juice are sugars, acids, vitamins, proteins, pectin, minerals, flavor, and aroma components. The composition of fruit juice depends on some factors such as variety, origin, processing procedure, and storage (De et al., 2009). At this point, using of nonthermal HHP method to process fruit juices is a promising way. Several researches have been conducted to identify the influence of HHP on nutritional features of fruit juices. Previous studies reported that HHP may cause a positive effect on berry juices in terms of their anthocyanin and flavanol contents (Patras et al., 2009a; Bodelon et al., 2013; Barba et al., 2012d). The effects of HHP (4000, 5000, 6000 bar; 900 s; 10°C, 30°C) and heat treatment (70°C, 120 s) on antioxidant capacity, cyanidin-3-glycoside,

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­ elargonidin-3-glucoside, ascorbic acid, and phenolics of blackberry p and strawberry purees were investigated in a recent study. There was no significant changes in ascorbic acid content of all pressure treatments (P > .05) whereas heat treated samples above the 120 s showed 21% ascorbic acid reduction when compared with untreated samples. HHPtreated samples indicated higher antioxidant capacity than those of heat treated ones. Cyanidin-3-glycoside and pelargonidin-3-glucoside contents of HHP-treated purees were not significantly changed compared to untreated samples, while the anthocyanin levels were decreased in heat treated samples. As a conclusion, researchers reported that HHP treatment is a promising technology to produce more nutritious berry purees (Patras et al., 2009a). Likewise Bodelon et al. (2013) indicated a constant remaining in ascorbic acid content and a well retention in anthocyanins of HHP processed strawberry puree at varied pressure levels (1000, 2000, 3000, 4000 bar) in comparison to untreated samples. The effect of HHP (2000, 4000, 6000, 8000 bar; 15 min) on strawberry juice anthocyanins has also been studied by Zabetakis et al. (2000). They carried out HHP treatment at 8000 bar and resulted it the most efficient way to preserve the anthocyanins in strawberry juice. In another study performed by Barba et al. (2012d) blueberry juices were subjected to HHP in the conditions of 6000 bar, 42°C, 5 min and some nutritional characteristics (antioxidant capacity, ascorbic acid, total phenolics, total anthocyanin content) were researched during preservation at 4°C. HHP exhibited better ascorbic acid level, phenolic content, anthocyanin content, and antioxidant capacity during storage in comparison with untreated samples. It was concluded that to preserve the bioactive components in blueberry juice pending storage, using high pressure could be a potential application. As it was previously reported by researchers, besides the parameters of HHP (i.e., pressure, temperature, and time), physicochemical features of berry juices such as pH and total soluble solid, and storage conditions (temperature and time) affect anthocyanin and flavonoid content (Cano et al., 1997; Altuner and Tokuşoğlu, 2013). Most studies have reported that vitamin C is not comparably influenced by HHP (Barrett and Lloyd, 2012). Barba et al. (2013) reported the influences of HHP treatment (2000, 4000, 6000 bar; 5, 9, 15 min) on anthocyanin stability, antioxidant capacity, vitamin C, and total phenolic contents as well as pH, brix, and color contents of blueberry juice. The total anthocyanin content is similar or increased when compared with the value predicted for the fresh juice. Antioxidant capacity of the samples at 4000 bar, 15 min and 6000 bar—all times showed the lowest contents (8%–16% reduction) in all treated and nontreated juices. More than 92% vitamin C was retained after all HHP treatments. However, HHP-treated blueberry juices at 2000 bar for all times

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showed an increase in total phenolic content. There were no significant differences in pH, brix, and color levels of the samples. Pomegranate has been considered as a therapeutic food due to its protective effects against some diseases (Vidal et al., 2003). Because of high bioactive compound content of pomegranate such as phenolics, flavonoids, anthocyanins, and minerals, consumers also demand to different products of this fruit produced as fruit juice, jam, and molasses, etc. for consuming all seasons (Lee et  al., 2005; Mirdehghan and Rahemi, 2007). Some quality properties of pomegranate juice processed by HHP (2000, 3000, 4000 bar; 5°C, 15°C, 25°C; 10 min) and conventional heat process (85°C, 10 min) were studied by Subasi and Alpas (2017). There were no significant decreases in antioxidant capacity, anthocyanin, and total phenolic contents (P > .05) of HHPtreated samples, whereas conventional heat treatment demonstrated an important decrease (P