Functionality of Bioactive Nutrients in Beverages

FUNCTIONALITY OF BIOACTIVE NUTRIENTS IN BEVERAGES

7

Rana Muhammad Aadil*,†, Ume Roobab*,†, Amna Sahar*,‡, Ubaid ur Rahman*, Anees Ahmed Khalil§ *

National Institute of Food Science and Technology, Faculty of Food, Nutrition and Home Sciences, University of Agriculture, Faisalabad, Pakistan, †School of Food Science and Engineering, South China University of Technology, Guangzhou, China, ‡Department of Food Engineering, Faculty of Agricultural Engineering and Technology, University of Agriculture, Faisalabad, Pakistan, § University Institute of Diet and Nutritional Sciences, Faculty of Allied Health Sciences, The University of Lahore, Lahore, Pakistan

7.1 Introduction Beverages cover all types of liquid foods that are purposely used for nourishment and quenching thirst. In fact, human being starts consuming beverages right after birth (breastfeeding) and until death (water). The classes of beverages other than alcohols, which may include fruit beverages (juices, squashes, nectars, syrups, etc.), stimulated beverages (tea, coffee, sports beverages, etc.) and functional beverages (dairy- and herbal-based beverages), or any mixture of different ingredients added in water that provide energy and nutrition. Fig. 7.1 explains the major classes of noncarbonated beverages marketed all over the world. These are not only the good source of energy, but also it provides a variety of nutrients like minerals, vitamins, and other bioactive components, that is, polyphenols, flavonoids, carotenoids, and bioactive peptides (BPs) that are necessary for the regulation of human body. Functional beverages like nutritional drinks support the old age community for meeting their nutritional requirement when they fail to complete it from their diet. These nutritional supportive drinks provide vitamins and minerals to help them for maintaining their nutritional need and proper body weight. Every type of beverage has its unique importance such as herbal beverages (ginger root, elderberry, hibiscus, cinnamon, fenugreek, bilberry, peppermint, onion, garlic, psyllium, goldenseal, licorice, cayenne pepper, etc.) having a variety of functional ingredients (gingerol, allicin, organosulfur compounds, arginine, lysine, tryptophan, threonine, etc.), which have antiinflammatory, a­ ntioxidative, Nutrients in Beverages. https://doi.org/10.1016/B978-0-12-816842-4.00007-1 © 2019 Elsevier Inc. All rights reserved.

237

Friut beverages

Fresh juices

Drinks

Calories

Calories

Minerals

Minerals

Vitamins

Syrup

Calories

Stimulating beverages

Squash

Calories

Nector

Tea

Coffee

Functional beverages

Sports drinks

Calories

Catechins

Caffeine

Minerals/ electrolytes

Vitamins

Theogallin

Flavonoids

Carbohydrates

Mineral

Flavanols

Carotenoids

Gallic acid

Quinic acid

Theanine

Methylxanthines

Minerals

Theafl avins

Thearubigins

Fig. 7.1  Beverage types and their functional ingredients.

Milk based

Bioactive peptides

Herbal drinks

Phenols

Flavonids

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Noncarbonated beverages

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antitumor, antidiabetic, anticholestrolemic, and antimicrobial characteristics. While in the case of stimulated drinks, tea has the greatest prevalent around the world, next to the water as tea or coffee have been consumed by billions of people from diverse cultures, different civilizations, and different regions of the world, at the starting of every day. These stimulating beverages have a variety of functional ingredients such as phenolic compounds and caffeine that improve cognitive function and promote health by reducing a number of chronic diseases risk such as cancer, cardiovascular diseases (CVDs), diabetes, etc. However, sports drinks continue to grow increasingly globally due to its advantages, it is provided in different games that provides energy (carbohydrates, amino acids, vitamins, electrolytes, etc.) to the players and these ingredients stimulate the mental and physical systems of the body that enhance the metabolic activity. Among beverages, fruit juices have a significant role in providing the bioactive ingredients like essential amino acids, carbohydrates, vitamins, minerals, and phenolic compounds together with its caloric contents, improve cardiovascular health and help to reduce the chronic inflammatory diseases. However, a variety of milk and milkbased beverages have been a part of daily dietary intakes that varies during the human life cycle. These dairy beverages have a number of nutritional and functionally bioactive ingredients such as BPs, which have distinct nutritional effects like gut functional agents, antihypertensive, antimicrobial, immunomodulatory, antioxidative, anticarcinogenic properties. The most important factor on the functionality of beverages is the losses of their bioactive compounds during processing (blanching and pasteurization), as well as some nonthermal treatment such as pulsed electric field (PEF), high pressure processing, ultraviolet radiation, etc. This chapter will give a detailed overview of noncarbonated beverages, their nutritional importance, and effect of processing treatment on their functionality.

7.2  Functional Drinks Containing Bioactive Components Functional beverages contain various bioactive components that could be the main reason for their functional and nutraceutical activities. Generally, these bioactive components belong to some common classes named as phenolic acids, flavonoids, and carotenoids. Detailed introduction of these main classes with their health implications has been discussed in this chapter (Table 7.1).

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Table 7.1  Summary of Plant-Based Bioactive Components and Their Functional Perspectives Common Name of Herb

Bioactive Components

Fenugreek

Flavonoids, Vitamin A, C, D, B1, B2, B3 Cinnamaldehyde

Cinnamon Bilberry

Garlic

Catechin, astragalin, quericitin, vitamin C Allicin, vitamin C, vitamin E

Functional Use

Reference

Hypoglycemic, antiulcerogenic, hypocholesterolemic, antiinflammatory

Singletary (2017)

Antibacterial, enhance insulin sensitivity, antitumor, antiinflammatory Antioxidant, antiinflammatory, gastroprotective, hypoglycemic

Hashemiravan and Ghahri (2017) Grohmann et al. (2017), Osada et al. (2017) He et al. (2007), Jayaraj and Lal (2017), Ugwu and Suru (2016) Attyah and Ismail (2017), Alexander and Williams (2016), Danwilai et al. (2017), Marx et al. (2017) Dyab et al. (2015), Shalayel et al. (2017)

Antioxidant, Antithrombotic, Antihypertensive, reduce serum cholesterol

Ginger

Gingerol

Hypolipidemic, Analgesic, antiinflammatory, hepatoprotective, antioxidant, immunomodulation

Peppermint

Eriocitrin, hesperidoside, rosmarinic acid Catechin, flavone glycoside, proanthocyanidins, Vitamin C Tannins, rutin, querecitin, vitamin C

Antispasmodic, carminative, antiallergic, antimicrobial, coolent, antioxidant

Cranberry

Raspberry leaves

Bacteiostatic, Antioxidant

Neto (2007), Peixoto et al. (2018)

Antidiarrhoel, antiinflammatory, astringent, regulate uterine contraction

Biel and Jaroszewska (2017)

7.2.1 Phenols Secondary plant products that have an aromatic ring carrying one or more hydroxyl groups which are phenolics. One of their major class is phenolic acids, which are located extensively in plant-based products in bound form (Ferreira et  al., 2017). Caffeic, coumaric, sinapic acid, and ferulic acids are the most known hydroxycinnamic acids (Aguilar-Hernández et  al., 2017) while protocatechuic, p-­hydroxybenzoic, ­syringic acid, and vanillic acids are hydroxybenzoic

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acids (De Beer et  al., 2017). Difference occurs in methoxylation and hydroxylation of aromatic rings of these derivatives. In foods, hydroxycinnamic acids are found as simple esters with glucose or quinic acid. Chlorogenic acid is combined with quinic and caffeic acids that is the most abundant soluble bound hydroxycinnamic acid (Matei, 2017). However, derivatives of hydroxybenzoic acid in foods are chiefly present in glucosides form (del Olmo et al., 2017) while glucose esters have been found merely that's why these are commercially available as synthetic sucrose esters and utilized as food additives or emulsifiers (Nagai et al., 2017). Dietary polyphenols (phenolic acids) are considered as powerful antioxidants and have been evaluated for the cancer treatments (Russo et  al., 2017). There in  vitro antioxidant activity is very high as compared to well-known antioxidant vitamins (De Ancos et al., 2017). However, the only mechanism through which polyphenols can exert their actions is antioxidation (Williamson, 2017). Other than antioxidation, polyphenols have different actions including antiviral, antibacterial, antimutagenic, antiinflammatory, antiproliferative, anticarcinogenic, and vasodilatory (Williamson, 2017; Ferreira et al., 2017; Russo et al., 2017). Several epidemiological studies have been reported that the relation between the intake of certain polyphenols can lower possible threat of many degenerative diseases, that is, chronic and CVDs. The intake of coffee can enhance high-density lipoproteins (HDL)-mediated cholesterol through its plasma phenolic acids from the macrophages, which proves its importance that coffee might have an antiatherogenic property (Teixeira et al., 2017; Murillo and Fernandez, 2017). It is proved that balanced diet can provide phenolic acids in abundance and plenty of them can be consumed daily by eating adequate amounts of fruits, vegetables, and whole grains. Mangoes, citrus fruits, berries, plums, apples, kiwis, cherries, onions, coffee, tea, red wine, whole wheat flour, oats, rice, and corn are the examples of foods having phenolic acids (Williamson, 2017; De Beer et al., 2017; Biegańska-Marecik et al., 2017).

7.2.2 Flavonoids Flavonoids are present in plants derived edible products, which are basically phytonutrients that often give color to foods. In human diet, the most abundant polyphenols are flavonoids, representing about 2/3 of all that are ingested. These are the products of secondary metabolism of plants like other phytochemicals. These can accumulate in cell vacuoles (water soluble) and frequently observed in the glycoside nature and sometimes as may be recognized as acyl glycosides, even though lower concentrations of methylated, acylated, and sulfate molecules are found (Kozlowska and Szostak-Wegierek, 2014). In human health, flavonoids play an imperative role. Animal-based (in  vivo)

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and in vitro studies have revealed that flavonoids have antimutagenic and antioxidant properties and may lower the risk of stroke and CVDs (van Dam et  al., 2013; Martinez-Perez et  al., 2014; Babu et  al., 2013; Ravishankar et al., 2013; Batra and Sharma, 2013). Iso-flavonoids, such as phytoestrogens, have many hormonal and nonhormonal functions in animals or in vitro, which recommended the consumption of these compounds for health benefits. Flavonoids have been proven for its antiproliferative functions to prevent tumor risk and growth, also known as antioxidants, which obstruct free radical to facilitate cytotoxicity and lipid peroxidation. Along with these properties, flavonoids are also recognized as weak estrogen agonists or antagonists, which is needed for the modulation of endogenous hormone performenses (Umar et  al., 2015; Fadaka et  al., 2015; Yasuda et al., 2017) as well as provide a defensive mechanism to combat chronic diseases including cancer and atherosclerosis. In addition, it provides an assistant in the regulation of menopausal signs (Correa et al., 2017; Fu et al., 2017; Sedighi et al., 2017; Mao et al., 2017). Consequently, these have been signified as semi-essential food components because of their enormous health advantages. Previous findings revealed the functionality of tea polyphenols associated with human health together with antimicrobial property, strengthening the capillary functions, and antioxidative nature that is capable of producing radioprotective effects (Liang et al., 2017; El-Desouky et al., 2017; Koech et al., 2013). Tea consumption increases the antioxidant capacity in human plasma that is linked with lowering risk of CVDs (Goszcz et al., 2017; Monteiro and Peluso, 2017). Interestingly, milk addition in tea can ­decrease or completely inhibit tea antioxidant properties due to milk-­ caseins interact with polyphenolic catechins from tea (Egert et  al., 2013; Ye et  al., 2013; Rashidinejad et  al., 2017). However, further investigation about the underlying biochemistry is required in this direction. Biological properties of flavonoids may account for cancer chemo prevention. Extensive considerations have been acknowledged in recent years to evaluate their capabilities for prevention of the cell proliferation, oxidative stress and cell cycle as well as to encourage apoptosis and detoxification of enzymes with activation of the immune system (Zhang et al., 2017; Hariri et al., 2017). Numerous procedures have been investigated for the evaluation of flavonoids imparting antineoplastic effects, such as antiproliferative, antiinflammatory, and antioxidative effects along with induction of detoxifying enzymes in conjugation with bioactivating enzymes inhibition (Asha et al., 2017; Semaan et al., 2017). Bananas contain anthocyanidins including cyanidin and delphinidin in ample amounts (Singh et al., 2016; Padam et al., 2014). Citrus fruits including lemons, grapefruit, limes, and oranges are high in

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f­lavanone (group of flavonoids) including naringenin, eriodictyol, and hesperetin (Selvamuthukumaran et al., 2017; Hwang et al., 2017; del Río et al., 2017). Apples, plums, pears, peaches, and apricots are members of the Rosacea family and are known to have high amount of epicatechin and catechin and are best if consumed with skin (Brahem et  al., 2017; Raudone et  al., 2017). Particularly, blue red and purple berries are high in flavonoids while darker and riper berries tend to have high flavonoid value and processing may decrease their flavonoid content. Cranberries and blueberries are containing high quantities of the flavanol group including myricetin and quercetin (Guo et al., 2017; Flores and del Castillo, 2015). Black grapes and blackberries are high in the flavonoids catechin and epicatechin while cherries, raspberries, and red grapes may be high in cyaniding and anthocyanidins (PeñaSanhueza et al., 2017).

7.2.3 Carotenoids Plant pigments that are responsible for their yellow, bright red, and orange color are carotenoids that play a key role in plant health. Therefore, consumption of foods that have carotenoids provide various health benefits to the human body. Different vegetables and fruits in which carotenoids are present: yams, carrots, sweet potatoes, watermelon, papaya, cantaloupe, spinach, mangos, kale, tomatoes, oranges, bell peppers, etc. There are over 600 known carotenoids in nature, found in plant cells, bacteria, and algae, and that belongs to a class of phytonutrients (“plant chemicals”) (Meléndez-Martínez et al., 2014). Following two are the main categories of carotenoids; carotenes and xanthophyll, both have different composition and molecular structure. Xanthophyll is the molecules known as hydrocarbons and found in orange and yellow fruits and vegetables, for example, pumpkin, cantaloupe, sweet potatoes, apricots and carrots. Association of word carrot with carotene will certainly relate the color of these pigments. Later class is oxygen-containing molecules, found in dark leafy greens such as kale, spinach, and broccoli (Al-Yafeai et al., 2018). Xanthophyll as well acts as an antioxidant and particularly have a great role in the health of human eyes (Grudzinski et al., 2017; Thomas and Harrison, 2016). However, carotenoids facilitate plants in photosynthesis by absorbing light energy (Solovchenko and Neverov, 2017). These also deactivate the free radicals, hence providing the antioxidant function as well as strong cancer-fighting characteristics (Soares et al., 2015). Vitamin A is crucial for vision, normal growth, and some carotenoids can be converted into this vitamin in the body. Furthermore, they also have antiinflammatory and immunity boosting properties and used to prevent CVDs (Yamagata, 2017; Amengual et al., 2017).

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7.2.4  Bioactive Peptides Milk has an ample variety of proteins that are vital for certain milk products manufacture and their characteristic nature, protects the body against enteropathogenesis. An array of bioactivities has been present in milk, which have a profound influence beyond nutrition. Within the protein molecules, peptides are dormant or inactive and they activated after releasing, during enzymatic digestion. These can be 2–20 amino acids per molecules are present in biologically functional peptides that are released from caseins and whey proteins (Mann et  al., 2017; Sánchez and Vázquez, 2017). These BPs named “Biopep” have a significant biological characteristic containing antioxidative, antihypertensive, opioid, antimicrobial, immunomodulatory and mineral binding actions, and anticytotoxic property (Nielsen et al., 2017; Singh et al., 2014). Most of the milk proteins have hidden, absent, or incomplete bioactivities, found in the original native protein. But after proteolytic digestion, these are release and activated as encrypted BPs of the original protein. Within the amino acid sequences of native milk protein, BPs have been identified. Proteolysis may release them during food processing or gastrointestinal transit. These bioactive compounds usually generate from digestive enzymes naturally present in milk, microbial enzymes, coagulants, and lactic acid starter bacteria (fermentation). During the processes of milk fermentation and cheese maturation, BPs enriched the dairy products after releasing from milk proteins (Aguilar-Toalá et al., 2017; Egger and Ménard, 2017). Numerous peptides derived from milk have demonstrated multifunctional features and particular peptide sequences that can perform different physiological activities. Different actions can be exerted by overlapping peptide sequences of the primary casein structure in certain regions. These fragments are considered as strategic regions, which are partially safe from more breakdown during proteolytic activity. A variety of peptide fragments have regulated various physiological actions while the peptides having different amino acid sequence, exhibited similar or different bioactive functions (Giacometti and Buretić-Tomljanović, 2017).

7.3  Functional Drinks Containing Herbal Extracts Herbs have been used for their medicinal and nutritional properties for centuries in the form of powder as well as extract. Variety of bioactive components has been reported in herbs in surplus amount that can easily be extracted and used in various drinks and food products for health benefits or value addition. Traditionally, in many countries, herbal extracts are still used for the prevention of many diseases like

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cancer, Alzheimer's disease, renal disorders, digestion problems (diarrhea, vomiting, cramps, and constipation), and diabetes (Koyama, 2017). Herbs have been reported to have properties like antiinflammatory, antioxidative, anticancer, antidiabetic, anticholestrolemic, and antimicrobial. Ginger root, elderberry, hibiscus, cinnamon, fenugreek, bilberry, peppermint, onion, garlic, psyllium, goldenseal, licorice, and cayenne pepper are the popular herbs and spices used for medicinal and nutritional purposes (Malongane et al., 2017; Panickar and Jewell, 2017; Violeta et al., 2017). Tea and its types (black, green, and oolong) used all over the world, are well known for their therapeutic values and energy activation properties. Camellia sinensis plant (leaves and buds) and its several breeds are used to prepare all types of tea. The presence of many valuable, bioactive, and volatile components makes it more appealing for almost every age groups (Chang et al., 2017). Tea along with other caffeinated beverages is being used for boosting up energy and nervous system and to relief tiredness. Most of the herbal extracts are used for the sake of value addition in these beverages. Tea and coffee itself contains catechin, caffeine, and other flavonoids that act as antioxidants in the body and play a vital role against free radicals and carcinogenic compounds and prevent from cancer and CVD (Koech et  al., 2013; Monteiro and Peluso, 2017). In south Asian countries, ginger is used in regular practice in tea for the purpose of making better taste and aroma, but bioactive components are leached out in tea and improve the nutritional parameters of tea (Malongane et al., 2017; Alexander and Williams, 2016). Almost all herbs contain different classes of flavonoids (responsible for plant coloration) especially flavone, flavanone, and flavanol. Flavonoids are important class of bioactive components that can easily be leach out in organic solvents but in aqueous medium, extraction is improved by mild heating. There are different means to extract flavonoids while used (traditionally) in hot beverages, herbal extracts are made in aqueous medium, when the purpose is to store the extract, organic solvents are used as a medium that is being evaporated after extraction and pure concentrated extract is achieved (Abdollah et al., 2017). From nutrition point of view, the best practice is to gain maximum benefits from herb (pure form). There are certain taste and flavor issues when herbal extracts are used in beverages due to some reasons that herbs contain intense taste and flavor. Generally, herbs used in beverages are ginger (Awe et al., 2013), garlic (He et al., 2007), peppermint (Dyab et al., 2015), onion (Han et al., 2000), capsicum (Nagasukeerthi et  al., 2017), turmeric (Singhal et  al., 2016; Pianpumepong et al., 2012), hibiscus (Monteiro et al., 2017a,b), fenugreek and licorice, cinnamon, clove, nutmeg, vanilla, and malt extracts beverage (Ghahri et al., 2017; Hashemiravan and Ghahri, 2017).

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Among these, ginger is a power house of well-known phenolic compound called as gingerol, which has a key role in performing a great number of functions inside the body after absorption. In beverages, ginger has been mostly used as fresh as in tea or in the form of extract in a minute quantity. Plenty of research work has proved in vitro as well as in vivo activities of ginger extract regarding important health issues starting from nausea, vomiting, weight loss, and chemotherapeutic effects (Marx et al., 2017; Attyah and Ismail, 2017; Danwilai et al., 2017). Garlic bulbs contain numerous bioactive components and amino acids such as allicin, organosulfur compounds, arginine, lysine, tryptophan, and threonine. Among these, allicin is the most valuable due to its functional and pharmacological importance (Ugwu and Suru, 2016). Allicin is formed when garlic is chewed or crushed and exposed to the enzymatic activity of alliinase, antioxidant, and antithrombotic, reduce cholesterol and immunoregulatory (Jayaraj and Lal, 2017).

7.4  Functional Components of Stimulated Drinks Stimulated beverages mainly include those drinks that accelerate the body functioning such as coffee and tea, or water-based artificial juice, which have series of components like taurine, ginseng, and guarana (sports drinks). The main effect of these components is on the central nervous system along with constriction of blood vessels that faster the heartbeats, increased respiration rate, and ultimately suppress the appetite. The main purpose of these drinks includes alertness, get rid of fatigues, stay awake and attentive, increase in motor and sensory activity, and boost up the energy levels.

7.4.1 Coffee Functionality Coffee is one of the most consumable product throughout the world, having important nutritional composition and evaluated for various health-related advantages such as alertness and boosting energy, inspiring mood, stimulating anxiety and insomnia, and relief from several syndromes such as headache, weakness, sedation, sleepiness, and irritability. Regarding to its bioactive composition, coffee contains about 2000 chemicals other than caffeine (the best-known component of the beverage) with some potential health impressions. The amount of these bioactive components varies according to the type of coffee beans, its brewing and preparation technique, and serving size of the beverage. Coffee consists of many functional ingredients like carbohydrates, lipids including diterpenes (cafestol and kahweol), proteins, nitrogenous compounds having amino acids, minerals especially

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potassium, caffeine, trigonelline, melanoidins, esters, and acids that consist of chlorogenic acids. Coffee is the most significant source of many dietary antioxidants such as phenolic compounds (chlorogenic, coumaric acids, caffeic, and ferulic). Themost important alkaloidcalled caffeine (1,3,7-­trimethylxanthine) appears as a viable antagonist on the adenosine receptors with a variety of physiological effects. This unique component provides a mild stimulation to the central nervous system, reducing sleepiness, ­fatigue, exhaustion, and drowsiness. On the other hand, it induces bronchodilation, stimulating respiration center in the respiratory system. Furthermore, it serves the other systems of the body such as cardiovascular system (increases heartbeat rate and the blood pressure) (Monteiro and Peluso, 2017), gastrointestinal system (gastrointestinal motility, enhanced gastric secretion, i.e., HCl and pepsin and biliary acids secretion), impart a thermogenic effect and accelerates lipolysis in the adipose tissue and urinary system (a mild diuresis).

7.4.2 Tea Functionality Tea (C. sinensis, Theaceae) leaves and bud’s product are the best widespread beverage all over the world. Tea can be categorized according to their cultivation and manufacturing such as unfermented tea is known as green tea, have fermented tea called oolong tea, and fully fermented is black tea while postfermented named pu-erh tea that is also recognized in some regions of the world. Other than its antibacterial and antiviral nature, tea is also known by its hepatoprotective and neuroprotective effects along with diuretic functions. It persuades restlessness, decreases the sensation of fatigue as well as reduces the risk of various chronic disorders like cancer, obesity, diabetes mellitus, atherosclerosis, or CVD. The main important components of green and black tea beverage include catechins, flavanols, theogallin, ascorbic acid, gallic acid, quinic acid, theanine, methylxanthines, minerals, volatiles, theaflavins, thearubigins, and caffeine. However, theanine, γ-aminobutyric acid, and major catechins (epicatechin gallate and epigallocatechin) are the active components of green tea other than vitamins, theobromine, theophylline, and gallic acids that lower the blood pressure and regulate the neural functioning. While oolong tea consists of monomeric catechins (8%–20%), theaflavins, and thearubigins, and numerous polyphenolic by-products (theasinensins and chaflosides). On the other hand, black tea has high antioxidative effect because it is manufactured from enzymatically oxidized fresh tea leaves.

7.4.3 Sports Drinks Functionality Intensive training is necessary for success in the game field, based on the performance to bare the training load without any injury or

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illness. At this stage, nutrition has to be considerably important that can be controlled by athletes to improve their performances. During events, it is difficult to take solid foods to fulfill their energy without interrupting the exercise, which can lead to dehydration that adversely effect on the mental and physical performance. That’s why, energy or sports drinks are available in the markets, which serve a variety of purposes including supply of substrate, electrolyte replacement, and prevention of dehydration when it is consumed during exercise. Furthermore, sports drinks can also be used before exercise for the hydration and energy recovery after the exercise. The formulation of sports drinks is very critical aspect, cannot rely on single formulation because of the fact that biological systems various among individuals in all the situations. Generally, sport drinks consist of carbohydrates (glucose, sucrose, oligosaccharides, and a blend of glucose and fructose) minerals, and electrolytes that enter the blood glucose pool during exercise and prevent hypoglycemic conditions. This phenomenon enhanced the exercise capacity by maintaining or uplifting the concentration of glucose circulates in the body. These ingredients of sports drinks provide characteristic taste, mouth feel, and palatability along with enhancing the need for water in the small intestine by sugars and sodium. They restore fluid, balanced the electrolytes, refilling of fuel stores, avoidance of illness, infection, and injury, psychological recovery, stimulation of the anabolic, and catabolic pathways that are involved in the adaptation process. However, the effectiveness of these drinks depends on the type and concentration of carbohydrates and electrolytes (sodium and potassium), osmolality, anion substances, buffer capacity, pH, and carbonation level. Active components include ergogenic compounds, that is, taurine, ginseng, aspartate, or glutamine, while other ingredients contain flavoring, vitamin, amino acids, herbal extracts, etc. Other important functions of sport drinks include muscle glycogen resynthesize (carbohydrate-based drinks), protein synthesis and tissue remodeling (milk-based drinks), brain effective skilled performance (sports drinks with neurotransmitter precursor amino acids or branched chain amino acids), and promoting hyperhydration (electrolyte and glycerol-based drinks).

7.5  Dairy Beverages Bioactive Components and Their Applications Milk being a complete diet possesses a huge range of bioactive compounds that perform appreciative functions on digestion. These components have a variety of nutritional and technological applications in different food products (Mohanty et al., 2016). A wide range of

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minerals, peptides, vitamins, enzymes, and lipids are among the list of bioactive components of milk, which are utilized to increase the nutritional value of foods. Milk proteins (whey and casein) have the highest bioavailability and derived peptides follow the same pattern. Milk proteins alone are very diversified in nature and variety of peptides can be derived from these proteins. These peptides can be classified based on functions as antimicrobial (Kamali Alamdari and Ehsani, 2017; Nielsen et al., 2017; Giacometti and Buretić-Tomljanović, 2017), antihypertensive and vasoactive (Fekete et al., 2013; Bhat et al., 2017b; Mazorra-Manzano et  al., 2017; Rai et  al., 2017; Elkhtab et  al., 2017), antithrombotic (Rafiq et  al., 2017; Tu et  al., 2017), hypocholesterolemia (Lucarini, 2017; Sgarbieri, 2017), antioxidative (Abdel-Hamid et  al., 2017; Sah et  al., 2018; Mada et  al., 2017), immunomodulatory (Hendricks et  al., 2017; Chalamaiah et  al., 2018; Reyes-Díaz et  al., 2017), and cytomodulatory peptides (Meisel and Fitzgerald, 2003). Dietary proteins not only provide energy but also the essential amino acids, which are key part of body functions including growth and maintenance. They are recognized by special components, enhancing the physicochemical and sensory properties of foods that are rich in proteins. Now, researchers are focusing on the functionally active proteins such as milk peptides, which have been potentially approved to regulate many processes in living systems, when these are activated by the gastrointestinal digestion or fermentation processes. These physiological functions are associated with the general health situation and their efficiencies are linked with the reduction of chronic diseases (Bhat et al., 2017b; Kamali Alamdari and Ehsani, 2017; Kang et al., 2017; Bouglé and Bouhallab, 2017) (Fig. 7.2).

7.5.1 Gastro-Modulatory Agents Dietary protein and peptides have been potentially used for the regulation of gastrointestinal functions including digestive enzymes regulation, intestinal tract management, control of nutrient absorption, and hydrolysis to amino acids (Sanchón et  al., 2018). There is an important peptide called casein-derived phosphorylated peptide or caseinophosphopeptides (CPPs), known for boosting calcium and magnesium absorption and vitamin D-independent bone calcification enhancement by restraining the calcium through distal ileum (Cao et al., 2017). Due to strong calcium binding affinity and activating voltage-dependent calcium channels, CCPs have been proved physiologically beneficial for the bone formation and prevention from osteoporosis, dental caries, hypertension, and anemia. These agents also enhance mucosal immunity by increasing the intestinal immunoglobulin production (Otani et  al., 2000). CCPs are also characterized as to activate the absorption of minerals (calcium, magnesium, phosphorus, iron, and zinc) and enhanced their bioavailability by

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Casein based peptides

GMP

CMP

Whey based peptides

CPP

Lactoferricin

Antimicrobial

Antihypertensive

Anticaryogenic

Anticarcinogenic

Immunomodulatory

Antimicrobial

Antimicrobial

Anticaryogenic

Anticaryogenic

Lactoferrampin

Antimicrobial

Gastromodulatory

Gastromodulatory

Fig. 7.2  Milk protein-based peptides and their health prospects.

providing binding sites for minerals. These are resistance to gastric digestion and help in mineral stability in intestines, serve as mineral carriers. These agents can be found in intestinal contents and stomach (distal small ileum) of adult humans after utilization of dairy product, that is, milk or yogurt (Mann et al., 2017). Other dairy peptides named glycomacropeptides (GMPs), caseinoglycopeptides, or nonglycosylated phosphorylated caseinomacropeptides (CMPs) establish the potential role in the regulation of intestinal functions such as slowing down the movement of stomach contractions in conjugation with inhibition of secretions. Stimulation and release of the satiety hormone cholecystokinin (CKK) is one of the main function of CMPs for controlling the food ingestion and digestion in the intestinal part of animals and humans, that is, duodenum (Alireza et al., 2017). CMPs have the ability to be used in functional foods as a prebiotic, which nourished healthy gut microflora without affecting food energy intake (Gustafson et al., 2001). In contrast to GMPs, CMPs are resistant to the intestinal enzymes so, it does not absorb in the intestine because during digestion process, intact CMPs have been investigated in the stomach (Fosset et al., 2002). β-Casein-derived opioid peptides called “β-casomorphins” that can also be a reliable source of gastro-modulatory agents, which absorb through gastrointestinal tract by affecting the smooth muscles, improve antisecretory function, and regulate the intestinal transport of electrolytes. These peptides have been detected in the duodenal

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chyme, plasma, and small intestine of newborn animal and human after the oral administration of casein or milk. On the other hand, after milk or yogurt ingestion, GMPs were absorbed through intestines and circulated into the blood, modulating the gut microflora and promote bifidobacterial growth (Cossais et al., 2017). CMPs and a-lactalbumin supplemented infant formula are available in the market that have same ability to promote the bifidobacterial activity as breast milk, whereas commercial products of GMPs are used for controlling the appetite and weight management (Brück et al., 2003).

7.5.2 Antihypertensive Agents Peptides derived from β-casein (isoleucine-proline-proline and v­aline-proline-proline) have been studied to act against the prevalence of hypertensive ability and, characterized as poten­ tially effective against CVD (Bhat et  al., 2017b). Generally, peptides associated with the prevention of CVD have the abilities to act as antihypertensive, antiatherosclerosis, antithrombotic, and hypocholesterolemia (Marcone et  al., 2017). Each of these peptides has its own mechanism of action and perform a different function but these are directly or indirectly associated with the prevention or cure of cardiovascular risk (Asledottir et  al., 2017). Angiotensin converting enzyme (ACE) inhibition, antioxidation, immunomodulation, opioid-induced blood pressure regulation, and endothelin-converting enzyme inhibition are among the mechanism that regulates the blood pressure and ultimately prevents the risk of CVD (Wu et al., 2017; Mann et al., 2017). ACE inhibitory peptides are the most important among all others because these have studied the most in human intervention studies, for example, lacto-tripeptides (Simonetti et al., 2017). But maximum efficiency is required by the absorbance of these peptides through small intestine and remains indigestible by the enzymes and hormones till reached the target site. These peptides need to stay in the blood and transport to the target site for their complete performance. Some of these peptides have been observed in monolayer-cultured human intestinal cells and proved to be resistant to the intestinal secretions. During enzymatic digestion of milk protein, several ACE inhibitory peptides have been released that further used in different food products for value addition and to improve the nutritional parameters. Whatever the mechanism, these peptides may follow, the important thing is their preventive activity against cardiovascular disorders and risks (Simonetti et al., 2017; Rai et al., 2017; Galland et al., 2017; Sultan et al., 2018). CMPs served as antithrombic agents by means of inhibited platelet aggregation that was induced through collagen and thrombin (Kamali Alamdari and Ehsani, 2017). However, fermented milk-based BPs

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(valyl-prolyl-proline and isoleucyl-prolyl-proline) along with hydrolyzed whey protein isolate (peptides) have been proved significantly effective to regulate the peripheral blood pressure thus exert antihypertensive effect in the body and prevent the risk of cardiovascular disorders. Commercially, these antihypertensive agents can be extracted from dairy-based products like milk, sour milk, cheese, and yogurt and available as casein hydrolysate drink, powdered fermented milk tablets, and casein hydrolysate tablets and capsules in the market (Bhat et al., 2017b).

7.5.3 Antimicrobial Agents Antimicrobial activity of the milk protein hydrolysates (peptides) has been identified (Kamali Alamdari and Ehsani, 2017; Nielsen et al., 2017; Delattin et al., 2017; Sah et al., 2018), which include CPPs, CMPs, GMPs, and whey-based peptide lactoferricin (derived from lactoferrin), lactoferrampin (Chen et al., 2017), and as1 and as2-casein (McCann et al., 2006). These dairy peptides are not in favor of the activity of various microorganisms (Gram-positive and Gram-negative bacteria) including Listeria, Salmonella, Escherichia coli, Helicobacter and Staphylococcus, fungi, and yeasts. Microorganisms such as viruses and bacteria as well as pathogens are facilitated by the carbohydrate fraction as they recognize carbohydrate receptors for adhering to the cells (Kamali Alamdari and Ehsani, 2017). CMPs interact with them inhibits the binding, and inhibits the growth of Streptococcus mutans, Porphyromonas gingivalis, and E. coli, protect cells from infection and provide antimicrobial services. On the other hand, lactoferricin antimicrobial activity depends on disruption of normal membrane permeability of the microorganisms (Chen et al., 2017), while lactoferrampin has been evaluated for candidacidal activity and sterile action against E. coli, Bacillus subtilis, and Pseudomonas aeruginosa (Kamali Alamdari and Ehsani, 2017). However, casein peptides (caseicin) have potential applications as dairy-based protectants against the growth and proliferation of neonatal Gram-negative pathogen Cronobacter sakazakii, Salmonella, and Klebsiella and the Gram-positive pathogen Staphylococcus aureus (Kamali Alamdari and Ehsani, 2017). These peptides (lactoferricin, CPPs, and GMPs) also play an effective role in maintaining oral health by anticariogenic properties, inhibiting to adhere cariogenic bacteria (S. mutans, Streptococcus sanguis, and Streptococcus sobrinus) with the oral cavity and providing favorable dental microbial composition. The anticariogenic effect of CPPs into salivary pellicles includes recalcification of tooth enamel, lowering the adhesion number of both S. sobrinus and S. mutans, whereas it is more favorable for rising of the Actinomyces viscosus

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population in conjugation with the modification of the dental plaque microbiota composition. Other than its adhesion prevention function, anticariogenic effect of GMPs based on the growth restrictions for the plaque-forming bacteria in oral mucosa, while lactoferrin has anticariogenic activity toward Gram-negative bacteria such as the dental cariogenic S. mutans. CPPs and/or GMPs have been utilizing in various commercial oral hygiene or dental care products, for example, mouthwash, toothpaste, and chewing gum.

7.5.4 Immunomodulatory Agents To regulate the immune functions, some casein and whey-based milk peptides and protein hydrolysates exhibited various activities such as antibody synthesis, regulation of cytokine, production of lymphocyte, stimulation of the phagocytic activities of macrophages, resistance to microbial infection by Klebsiella pneumoniae, alleviate allergic reactions, and enhance mucosal immunity in the gastrointestinal tract. Dietary CMPs play a vital role in maintaining immune system by the production of IgG antibodies inducing the proliferation of spleen lymphocytes through inhibition of mitogens, depending on the availability of sialic acid. This suppressing activity helped newborn mammals to respond environmental antigens and regulate their immune system. CMP can also be used for the manufacturing of immunosuppressive foods while as1-casein-derived peptide commercially used as ingredient in the preparation of various food items like soft drinks and confectionary foods. Casein-derived peptides have been investigated for stress-relieving properties as they act like anxiolytics. Other than CMPs, immunomodulatory effect has been identified for opioid peptides (hydrolyzed casein fractions) (Bedini and Spampinato, 2017; Fernández-Tomé et  al., 2016; Trivedi et  al., 2016; Stefanucci et al., 2018; Garg et al., 2016). Opioid peptides have agonistic or antagonistic activities and can bind with opioid receptor that can be found in gastrointestinal tract of mammals, nervous, endocrine, and immune systems. On the other hand, GMP and its derivatives have also been investigated for immunomodulatory functions including immunosuppressive effects on the production of immunoglobulin antibodies, and immunoenhancing effects for human macrophage proliferation along with its phagocytic activities improvements. CPPs cytomodulatory peptides prevent the growth of cancerous cell and regulate the activity of neonatal intestinal cells or immunocompetent cells to maintain healthy immune system. Moreover, peptides released during the process of milk fermentation have been evaluated for antitumor properties and cytomodulatory or immunomodulatory functioning. Inhibition of tumor expansion was seen

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through modulation of host’s immune response by peptides resulting from bacterial proteolysis (Chalamaiah et al., 2018). Commercial infant formals have been utilizing whey-based peptides that released during digestion.

7.5.5 Antioxidative Agents In dairy beverages, bioactive compounds have many other functions such as antimicrobial, antithrombotic, antihypertensive, opioid, immunomodulatory, related to the gastrointestinal system, cardiovascular system, nervous system, and immune system, while antioxidant effect is one of them. Dairy peptides are derived from casein (Cao et al., 2017; Fernández-Tomé et al., 2016) and whey proteins (Moura et  al., 2017). Antioxidative peptides bind free radicals and metal ions, conversion of cysteine to glutathione that protect the cell from enzymatic and nonenzymatic lipid peroxidation ultimately inhibit oxidative cell damage (Sánchez and Vázquez, 2017). Antioxidative properties of peptides developed by hydrolysis of casein under action of digestive enzymes in fermented milk. These peptides were derived from amino acids (cysteine, methionine, tyrosine, tryptophan, lysine, and histamine) and caseins, have been utilized in different food stuff for the prevention of oxidation, and proteolysis due to high hydrophobic nature (Mann et al., 2017). These peptides have proven to be safe and healthy compounds that are cheap, highly active, having low molecular weight and easily absorbed with no hazardous immunoreaction, exhibited nutritional and functional properties. These peptides including β-lactoglobulin peptide inhibit lipid oxidation by their scavenging properties for free radicals and transit metal ions and preventing oxidative stress. Antioxidative activity depends on different factors such as composition, structure, peptide sequence, and hydrophobicity, while the redox potential enhancing the radical-scavenging properties of peptides. However, operational conditions like structure and concentration of peptide, type of protease, and degree of hydrolysis may affect the antioxidant potential of protein isolates. Antioxidative peptides can produce from microbial fermentation and through the action of digestive enzymes. However, enzymatic hydrolysis of β-conglycinin and glycinin leads to increased antioxidant activity by the development of hydrophobicity and decreases allergenic agents. Furthermore, enzymatic hydrolysis releases free amino and carboxyl groups by breaking down peptides, enhancing its antioxidative property. On the other hand, fermentation can result in enhancing the nutritional value of foods, because of the releasing of BPs form proteins by microbial proteases. Antioxidative peptides are effective in the prevention of various degenerative diseases such as type II

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diabetes (by impairing the secretion of insulin or enhancing resistance against insulin) and metabolic syndrome related to cardiovascular complications include alteration in metabolism of lipids. These diseases rely on oxidative stress, which when controlled by antioxidative peptides may prevent proliferation and complication of the diseases.

7.5.6 Anticarcinogenic Agents Different strategies have been evaluated to use these bioactive components as anticarcinogenic peptides, which are associated with chemopreventive properties, immunomodulatory activities, and reduction of tumor proliferation as well as apoptosis. The food-derived BPs have been potentially used for the cancer treatments or auxiliary therapy of cancer, providing a substitute to highly expensive and adverse consequences of chemotherapy, radiation, and surgery. These bioactive components block, reverse, or retard the process of mutation, abnormalities, and carcinogenesis. These alternative anticarcinogenic molecules have the high affinities for the negatively charged structures of cancer cells, readily absorbed in cell matrix, target specific, and have less side effects or toxicity along with other properties (low price, high bioavailability, and effective for all stages of cancer), made them easily accepted by the consumers (Hernández-Ledesma and Hsieh, 2017). The cytotoxic milk peptide, that is, lactoferricin, against cancer cells has been derived from lactoferrin (whey proteins) (Kanwar and Kanwar, 2013). Breast cancer treatment based on the utilization of lactoferrin have the ability to inhibit growth of cancer cells by suppressing signaling of nasopharyngeal carcinoma cells (Deng et al., 2013). These have been proved significantly for the reduction of breast cancer (Sun et al., 2012), lung cancer treatments (Tung et al., 2013), colon, fibrosarcoma, leukemia, oral, and ovarian cancer cells, without harming normal lymphocytes, fibroblasts, endothelial, or epithelial cells. Another chemopreventive peptide (whey protein-derived α-lactalbumin combined with oleic acid) called bovine alpha-lactalbumin also have been effective for the chromatin condensation, modulation of carcinogenesis biomarkers, and cell shrinkage ultimately cancer cell death. On the other hand, antiproliferative peptides (opioid casein protein derived) β-casomorphin have been demonstrated for an effective tool of preventing colon cancer (Brinkmann et al., 2013; Pepe et al., 2013).

7.6  Effect of Thermal/Nonthermal Processing in Bioactive Nutrients Present in Functional Drinks Plant-based beverages contain a substantial number of bioactive components that have many functional roles in the body if consumed

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as fresh, but processing may have some effects on these components. These bioactive components may include carotenoids and retinols, tocopherols, phenolic compounds, flavonoids, sulfur-containing bioactive molecules, and folates. Domestic thermal processing is reported to have worst effects on bioactive nutrients due to their heat-sensitive nature. Although plant-based beverages provide good nutrition even after thermal processing, but it can be more valuable if they are processed nonthermally to provide better nutrition in the form of natural functional nutrients (Aadil et al., 2015a). Moreover, it is nowadays the demand of consumers to purchase nutritional-rich food products rather than energy-rich food products as these are reported to possess health problems like obesity and hypercholesterolemia (Lee et al., 2017).

7.6.1 Thermal Processing Bioactive nutrients need to be retained in functional drinks to play their role as functional nutrient, so processing conditions must be modified in such a way that the loss should be minimum. Domestically, beverages are processed thermally, and different thermal treatments are applied as per requirement of the beverage. Some bioactive nutrients can be incorporated and retained during domestic processing, but the problem is that most of bioactive nutrients are heat sensitive and their retention in beverages demand modified processing conditions in order to confirm their presence in final product (Petruzzi et  al., 2017). To manage these issues, novel processing technologies are need of the time to adopt them in processing lines for manufacturing of new and beneficial functional beverages. Most of the fruit juices available in the market are processed by domestic thermal processing and only provide nutrition in the form of calories from carbohydrates or through fortification and enrichment of some minerals and vitamins (Fig. 7.3).

7.6.1.1  Effect of Blanching Blanching is a technique that is used to inactivate such enzyme that may harm the nutritional components of fruit products. Different methods were introduced nowadays to blanch food products including water blanching, steam blanching, in-can, and vacuum-steam blanching. Among these, water blanching that ranges from 75°C to 95°C for 1–10 min due to low running cost. Blanching is basically performed as a pretreatment for many operations like pasteurization, freezing, and drying to make the food product more stable as enzyme has been inactivated. It can inhibit or reduce the thermal degradation by tissue disruption and releasing of anthocyanins from the vacuoles or other cellular structures ultimately improves the extractability and stability in the treated product (Nowacka et al., 2018; Deylami et al.,

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Thermal treatemtents

Blanching

Non thermal treatments

Pasturization PEF

Ultrasonication

Ozonation

Irradiation

Sterlization

Fig. 7.3  An overview of thermal and nonthermal processing techniques for beverages.

2016). Blanching has increased the level of radical-scavenging activity of the juice by the retention of phenolic components. Significant increase in anthocyanin and cinnamate content was seen in blueberry juice (Rossi et al., 2003) and increased retention of phenolic components in bottle gourd juice after blanching for 80°C and 4 min ohmic blanching (Bhat et  al., 2017a). Effectiveness of blanching process depends on its type (conventional blanching and ohmic blanching), boiling temperature and treatment time, blanched medium (water or steam) (Nowacka et al., 2018; Guida et al., 2013), and surface area of the treated product. Even the temperature is kept low in blanching, but some losses have been reported in heat labile bioactive components such as anthocyanins (Kidon and Czapski, 2007) and vitamin C (Skrovankova et al., 2015). Anthocyanins are considered as most heat-sensitive bioactive components as Pacheco-Palencia et al. (2009) have observed the loss of anthocyanin content in acai fruit when heated for 1–60 min at 80°C whereas other phytochemicals remain unaffected at this temperature while blanching at 95°C for 3 min in combination with pasteurization during processing of Aronia into juice resulted in significant loss of anthocyanins (Wilkes et al., 2013). Vitamin C (ascorbic acid) is another heat-sensitive bioactive component that the loss was occurred during blanching of fruits. The loss of ascorbic acid during blanching might be due to the breaking down of the dehydroascorbic acid (DHAA) and then dehydroxyascorbic acid degradation. However, novel blanching techniques such as the combination with high-pressure processing, ultrasound, microwave blanching, infrared blanching, and radiofrequency blanching have been evaluated for improving the functionality of food and beverages (Reis, 2017).

DPCD

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7.6.1.2  Effect of Pasteurization The heat labile characteristics or process severity led to thermal degradation that is increased for liquid foods (more prone to degradation) due to their unsaturated structure chemical structure. For this reason, sometimes degassing is done as a pretreatment before thermal processing that brings a considerable improvement in the retention levels of bioactive compounds. For avoiding losses of valuable components during pressing, there should be a complete knowledge of product’s chemical nature and type of the final product, as well as its effects on human health should be evaluated. Pasteurization is a basic step of processing of fruit products especially juices to make them free from microorganisms and some destructive enzymes like, polyphenol oxidase (PPO), pectin methyl esterase (PME), polygalacturonase (PG), peroxidase (POD), and others. Pasteurized fruit juices have longer shelf life due to inactivation of these enzymes and the absence of putrefactive microorganisms, but it imparts some negative effects on nutritional profile of juices, that is, loss of vital bioactive components. Studies proved that some heat labile bioactive components like vitamin A, total carotenoids, polyphenols, and vitamin C in mango puree may be affected by the severity of pasteurization which have been reported earlier in durian juice, mulberry fruit extract, pineapple juice while ascorbic acid and carotene in canned mango, papaya, and litchis (Beyers et al., 1979), quercetin, kaempferol, and myricetin in industrially processed acerola, cashew apple, pitanga juice, and pulp (Hoffmann-Ribani et al., 2009). Thermal treatment also led to red colored lycopene pigments degradation in tomatoes, which can cause nonenzymatic browning or Maillard reaction because of severe processing conditions (Jayathunge et al., 2015). However, there is an important aspect for degradation of bioactive components as it depends on the severity of the pasteurization treatment such as ascorbic acid in degassed tamarillo nectar remained stable up to the treatment temperatures, that is, 80–95°C for 10 min. Moreover, the same study was evaluated for nondegassed samples of tamarillo nectar, which showed a significant decrease in bioactive components (vitamin C and DHAA), ultimately degraded totally along with degradation in carotenoids (5,8-epoxidation and cis-isomerization) while total carotenoid content almost remains unaffected. These studies proved that the reduction of bioactive components was strongly related to its treatment parameters and pretreatments such as time, temperature, product type, degassing, etc. (Mertz et al., 2010). Among these bioactive nutrients, vitamins are among the most sensitive to thermal treatments (especially vitamin C), which is degraded by a chain mechanism of oxidation due to harsh sterilized treatments. Its degradation mechanism depending on different factors such as

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processing type, aerobic or anaerobic pathway, and catalyzed or uncatalyzed pathway. However, result of aerobic oxidation will lead to intermediate products such as DHAA, that is, further converted into 2, 3-diketogulonic acid (DKGA) while anaerobic oxidation produces intermediate keto-ascorbic acid and its anion keto-monoanion ascorbic acid during ketonization. However, these degradation forms further converted into delactonization forms DKGA. Other important bioactive components include carotenes and xanthophylls present in fruit beverages, which are also sensitive to heat, oxygen, and light (may undergo isomerization) and thermal pasteurization may affect their stability. Their degradation was evaluated in cashew apple juice during heat treatment of 60°C and 90°C for 1, 2, and 4 h in a water bath and mechanisms involved in their degradation were isomerization, epoxidation, and cleavage (Zepka and Mercadante, 2009). Elevation in the levels of oxidative products led to the reduction of bioactive components (carotenes, xanthophyll, lutein, and violaxanthin), which is catalyzed by increase in time and temperature. During thermal processing, the main reason of degradation in antioxidants is isomerization and oxidation. Polyphenolic compounds such as p-hydroxybenzoic, protocatechuic, ferulic, vanillic, syringic acid, pelargonidin, peonidin, cyanidin, and their glucosides have some considerable influence on the functionality of fruit-based beverages. Although, these are not significantly affected by oxygen during heating, but anthocyanin degradation (along with the formation of oxidative product chalcone) was observed (80°C for 60 min) Euterpe precatoria species (Sadilova et al., 2007). Rate of anthocyanin degradation was shown to be directly related to heating method and exposure times, but it was also confirmed that the reason for indirect oxidation was phenolic quinones generated by PPO and POD (Skrede et al., 2000). Indirect oxidation rate was influenced by anthocyanin's structure (e.g., non-o-diphenolic anthocyanin degraded slowly while o-diphenolic anthocyanin rapidly degraded through oxidation) and secondary oxidation products (Sarni-Manchado et al., 1997).

7.6.2 Nonthermal Treatments Every processing operation has a common purpose that is to make the beverage shelf stable as natural beverages have low shelf stability. On the other hand, most of thermal processing technologies have been designed to make heat-sensitive bioactive components available in the beverages other than improving shelf life of the beverage. While nonthermal processing technologies are a good practice to make the beverages microbiologically stable and fresh for comparatively more time than thermal technologies as well as to produce nutritionally better final product (Roobab et  al., 2018). Modern nonthermal

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t­ echnologies in juice and beverage processing include high-pressure treatment (Varela-Santos et al., 2012; Aadil et al., 2017), ultrasonication (Aadil et al., 2013, 2015a; Abid et al., 2014; de São José et al., 2014), PEF treatment (Pal, 2017; Aadil et al., 2015b), and their combination (Aadil et al., 2018a,b; Zhu et al., 2017), whereas traditionally juices and beverages are being processed by low pasteurization and high pasteurization treatments. These traditional treatments are being aided by instant freezing for better flavor retention and for packaging purpose (Leposavić et al., 2016) (Fig. 7.3).

7.6.2.1  Effect of PEF PEF is one of the most extensively studied nonthermal technology that have been applied on food and beverages for microorganism inactivation as well as retention for most of the bioactive components by leading to enhance extraction yields of metabolites (JiménezSánchez et  al., 2017). Researchers evaluated fruits bioactive components and demonstrated that PEF works better as compared to the conventional treatments in the case of functional components especially vitamin C because it is sensitive in nature and can be easily destroyed by the harsh treatment conditions (Yilmaz and Evrendilek, 2017). While some studies determined the impact of PEF treatments on bioavailability and accessibility of functional ingredients as well as their effect on human well-being (Rodríguez-Roque et  al., 2015). Besides, there is insufficient researches in terms of PEF influence and benefits on human health. However, PEF’s effectiveness depends on the treatment time and intensity such as shorter exposure to high-­ intensity pulsed electric field (HIPEF) resulted significantly to higher retention of ascorbic acid as compared to extended HIPEF treatment duration. Moreover, PEF was reported for higher vitamin C retention, immediately after this treatment as well as during the storage of soymilk beverage (Morales-de La Peña et al., 2010a,b) and orange juices (20 kV/cm pulses at 40–50°C) (Buckow et al., 2013). These observations were identical to the findings of OdriozolaSerrano et al. (2009), who processed the strawberry juice at 35 kV/cm of constant field strength for 1000 μs whereas bioactive components like vitamin C and anthocyanins were found at their maximum antioxidant activity at 232 Hz frequency for 1-μs pulse width. Their relative retention was observed to be in the range of 87%–102% under all experimental conditions, which was higher than the reported quantity. Antioxidant ability of a mixture of pineapple, kiwi, orange juice, and soy milk was checked, and PEF did not pose any effect, irrespective to the total time for treatment (800 or 1400 μs) and it was different from thermally processed (90°C, 60 s). In addition, during storage of 60 days, antioxidant potential of thermally treated product was shown to be decreased while the sample treated with PEF showed higher

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a­ ntioxidant potential (Morales-De La Peña et al., 2010a,b). Moreover, this technology has been reported to be used as a pretreatment for better extraction yield of several fruits and vegetables (e.g., apple, sugar beet, grapes, and carrot) (Corrales et al., 2008). The effect of PEF processing was comprehensively studied in the bioactive compounds of fruit juices (Yilmaz and Evrendilek, 2017). In HIPEF-processed watermelon juice that holds up to 87.6%–121.2% lycopene, which is over the extent of processing parameters. The possible mechanism for increase in lycopene content is that PEF induces the cell permeabilization and intracellular pigments to be released. It can be imagined that such increase in the intensity of electric field could make the watermelon cells in stress trigger, resulting in the production of lycopene as a secondary metabolite, which can accumulate and stimulate metabolic activity. On the other hand, the overall antioxidant ability of watermelon juice was affected as an increase in the pulse width and inverse effect was seen between the antioxidant activity and the pulse width. Research has precisely proven the amount of PEF processing for the preservation and maximizing secondary metabolite extraction that ideally should be in range of 1–10 kV/cm. Therefore, lower the field strength generally higher will be the juice extraction as well as secondary metabolites from food medium. In contrast, some studies showed bioactive component degradation after PEF treatment, due to the reason that it may induce some chemical reactions, may not completely inactivate enzymes like POD, β-glucosidase, and POD and direct impact of the treatment on functional components. That effect has catalyzed the oxidative reactions and may trigger bioactive losses along with some changes in the pH, temperature, and chemical composition of the food and beverages (Rodaite-Riseviciene et  al., 2014). On the other hand, β-glucosidase activity of strawberry juice has been reported to increase during PEF treatment of 35 kV/cm at 50 Hz for 1000 μs (Aguiló-Aguayo et al., 2008). Moreover, another PEF treatment carried out at 35 kV/cm for 800 and 1400 μs at 200 Hz had no effect on the total phenolic content of a blended juice of orange, pineapple, kiwi, and soymilk. However, considerable reductions were reported for the concentration of vitamin C that resulted in the reduction of antioxidant activity of the product (Morales-De La Peña et al., 2010a,b).

7.6.2.2  Effect of Ultrasonication Ultrasonication is one of the most reliable nonthermal treatment that has been extensity evaluated with promising results for reduction in bioactive components in food and beverages. But, it can also degrade some of the polar functional components due to the chemical reactions that was going through the effects of cavitation m ­ echanism

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effect called pyrolysis that start the production of entrapped gas bubbles in the liquid medium that may be free flowing. For example, ascorbic acid degradation has been demonstrated by the formation of free radical during sonication. The effect of hydroxyl radical formation has been catalyzed by degassing of the food as it could increase the dissolving level of O2 and N2 in liquid food by filling of sonication with water vapor and gases (Tiwari et al., 2009a). Ascorbic acid degradation involves the interactions between free radicals and ascorbic acid, thermolysis, which generates −OH radicals by the Maillard reaction that is the main reason to oxidizes the bioactive components like vitamin C and total phenols (Czechowska-Biskup et al., 2005). In the same way, ultrasound caused the degradation of lycopene, as a result of the formation of hydroxyl radicals that have been produced by acoustic cavitation in food extracts (Pingret et al., 2013). However, that reaction has been triggered by the addition of a small amount of water, which can decompose lycopene into a lipophilic ­bioactive antioxidant or isomerization of lycopene, decrease of lycopene bio-­accessibility (Anese et  al., 2013), whereas in the case of watermelon juice some ­oxidation compounds such as methyl-heptenone, laevunilic aldehyde ­acetone, and glyoxal have been observed (Rawson et al., 2011).

7.6.2.3  Effect of Ozonation Due to great biocidal effectiveness and extensive antimicrobial nature, the interest in ozone as a preservation technology was developed. The food industry uses the ozone treatment in the gaseous form for cleaning and disinfecting of fruits and vegetables for retaining quality during storage (Aadil et  al., 2018a,b). Concerns in prospective of ozonation process for liquid food were developed after the FDA approved ozone to be used as a significant treatment in the food industry (Cullen et al., 2009). Ozone has various potential applications as antimicrobial agent if it is compared with the conventional antimicrobial agents such as chlorine, potassium sorbates, etc. Concerns have been made for the application of ozone as a disinfectant that can be used for prolonging storage of various exotic fruits or their products (Brodowska et al., 2017; Pandiselvam et  al., 2017; Tai and Martin, 2017; Okpala, 2017; Brié et  al., 2018; Tzortzakis and Chrysargyris, 2017) Generally, ozonation (level of 0.15–5.0 ppm) has been exposed for growth restriction of the spoilage bacteria and yeasts. Barboni et al. (2010) compared the effect of ozonated storage with air storage for 7 months and total ascorbic acid content of kiwi fruit was checked. In the chamber, concentration of 4 mg/h ozone at 0°C and humidity of 90%–95% was applied and no significant change was observed. Pérez et  al. (1999) have investigated efficiency of ozone

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treatment of 0.35 ppm at 2°C on the postharvest quality of strawberry. The arbitrary in strawberries, the increase in ascorbic acid levels has resulted in response to ozone exposure. However, Ali et al. (2014) observed a significant increase in ascorbic acid (28.4%), total phenolic (14.3%), beta-carotene (82.2%), and lycopene (52.8%) contents and enhanced antioxidant activity (21.9%) in papaya after ozonation at 3.5 ppm for 96 h at 25°C. Similar effect was investigated for kiwifruit (total carotenoid increased to 2.1%, 2.5%, 2.8%, and 6.8% according to the exposure time of 8, 24, 72, and 144 h with 0.3 ppm ozonation, respectively) (Minas et  al., 2010), tomato (phenolic compounds was increased to 50% with 10 ppm, 10 min, 20°C) (Rodoni et  al., 2009), pineapple and banana (polyphenol and flavonoid content increased to 15.7% and 32% in pineapple and 8.2% and 14.7%, respectively, in banana) (Alothman et al., 2010; Du et al., 2016). On the basis of these studies, it can be concluded that ozone treatment may increase the antioxidant potential of various fruits but at the same time it reduces the ascorbic acid content that may affect its efficacy. Ozonation also resulted to have negligible influence on anthocyanin levels of strawberries (Lozowicka et  al., 2016) as well as of blackberries (Barth et  al., 1995), while an increase in total phenolic components and decrease in the levels of anthocyanins have been observed in fresh-cut papaya (Yeoh et al., 2014) and blackberry (Tiwari et al., 2009b). Sensory qualities of food may be affected, if high doses of ozonation are applied, other than the quantity that is enough for proper decontamination. In some cases, ozonation may promote spoilage in foods by oxidation, so it is not universally beneficial. Due to excessive use of ozonation, some quality defect could occur including discoloration, surface oxidation, which developed undesirable odors (Gertzou et  al., 2017). Studies have proven that the consequences of ozonation on physiological qualities of fruits depend on the time and ozone dose, application type, and chemical composition of food (Pandiselvam et al., 2017; Cullen et al., 2009). Conflicting researches have been observed in the literature related to ozonation and ascorbic acid contents of the food. The potency of ozonation on total phenol, vitamin C, and flavonoid components present in different fruits like pineapple, guava, and banana investigated by Alothman et al. (2010), which were exposed to ozone at a flow rate of 8 mL/s for 30 min. In pineapple and banana, total phenol and flavonoid contents were increased considerably when exposed to ozone with a simultaneous increase in antioxidant contents and 1,1-diphenyl-2-picrylhydrazyl values for up to 20 min, whereas in guava, the treatment effect was the opposite. However, results have proven that ozonation considerably reduces the vitamin C levels in all of these fruits while the study for effect of ozonation on other functional compounds of exotic fruits

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is limited. However, because of its intensive oxidizing property, ozonation is expected to be the major cause of the important phytochemicals and antioxidant bioactive compounds losses. During ozonation of the fruits, the main reason that could have involved in the degradation of ascorbic acid is it may be the stimulation of ascorbate oxidase enzyme which is activated under stress condition like chemical exposure (Alothman et al., 2010).

7.6.2.4  Other Treatments A nonthermal treatment called dense-phase carbon dioxide (DPCD), vitamin C and β-carotene losses have been observed at 8–35 MPa for 5–60 min at 35–65°C, respectively, in melon juice. Although, ascorbic acid has the higher stability in low pH environment of liquid foods, as a result of CO2 treatment, but its oxidation catalyzed in oxygenated environment (Chen et  al., 2010). However, DPCD processing is better substitute of pasteurization (90°C for 60 s) or thermal treatments for more retention of bioactive components in melon juices (ascorbic acid content was 6.4 times increased at 35 MPa, 55°C for 60 min) (Chen et al., 2010), enhanced anthocyanin stability in grape juice (34.5 MPa, 8% and 16% CO2, 30°C for 6.25 min) (Del Pozo-Insfran et al., 2006), increased antioxidant capacity (ascorbic acid contents) in guava puree, which was treated at 30.6 MPa, 35°C for 6.8 min (Plaza et al., 2010), and antioxidant activity increased in apple juice at 15 MPa, 35°C for 15 min (Porto et al., 2010). Similarly, gamma irradiation within the range of 300 and 600 Gy increased the amounts of phenolic content and antioxidant activity in apple (Mostafavi et al., 2012) as well as ultraviolet treatment for carrots with UV-B (1.3–12 kJ/m2) (Du et al., 2012), tomato with UV-B (6.08 kJ/m2 for 60 min) (Castagna et al., 2014), and pineapple, banana, and guava with UV-C (2.158 J/m2 for 30 min) (Alothman et al., 2009). In contrast, electron beam irradiation (1–3.1 kGy) has been observed in mango with no changes in total phenolics and carotenoids but increased flavanol levels (Reyes and CisnerosZevallos, 2007), whereas gamma rays (500 Gy) or UV-C (30 min) reduced phenolic and antioxidant activity during storage of mangoes (Chatha et al., 2013).

7.7  Conclusion and Future Trends Functional beverages impart various health implications upon the consumption due to the presence of various bioactive components, that is, phenols, flavonoids, carotenoids, peptides, vitamins, and minerals, but prior to their activity. These compounds need to be absorbed by the body due to these components, which have very low

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­ ioavailability and less absorbed by the body. As far as, the absorbance b of bioactive component is concern, it can be increased by developing such processing techniques that can the maximum retention. Loss of these compounds during processing is another concern that may degrade the value of functional beverages. Generally, these are proved to be sensitive against thermal processing and either degraded or destroyed during processing. So, keeping in mind these concerns, there is need to use such methods in processing of beverages that mainly focus on the minimum loss of these components so that the consumers may take maximum benefits from functional beverages. Use of nonthermal techniques like pulse electric field, ultrasonication, and ozone treatment would be a better solution for these concerns, because these techniques have been verified for microbial safety as well as shelf stability of beverages.

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Further Reading Guderjan, M., Töpfl, S., Angersbach, A., Knorr, D., 2005. Impact of pulsed electric field treatment on the recovery and quality of plant oils. J. Food Eng. 67, 281–287.