Role of Encapsulation in Functional Beverages

ROLE OF ENCAPSULATION IN FUNCTIONAL BEVERAGES

6

T. Ozdal*, P. Yolci-Omeroglu†, E.C. Tamer† ⁎

Department of Food Engineering, Faculty of Engineering, Istanbul Okan University, Istanbul, Turkey, †Department of Food Engineering, Faculty of Agriculture, Bursa Uludag University, Bursa, Turkey

6.1 Introduction There is a growing demand for health improvement or disease prevention through the incorporation of bioactive compounds including polphenols, vitamins, minerals, omega-3-fatty acids, bioactive proteins or peptides, and probiotics into functional beverages and also preventing them from processing and storage conditions to improve their bioavailability. They have poor stability in processing and storage conditions and their poor bioavailability or chemical variability when subjected to the conditions of the upper gastrointestinal tract (GIT) significantly decrease their related health benefits. Encapsulation is an appealing method to entrap bioactive compounds within a polymer material for the preservation and delivery of bioactive compounds at the right time and to a targeted site (Ezhilarasi et al., 2013). The most important criteria to decide the selection of an encapsulation material are functionality provided to the final product, potential limitations of coating material, concentration of encapsulates, stability needs, type of release, and economic limitations. The materials used for the fabrication of protective shell of encapsulates must be biodegradable, food grade, and must form a barrier between the internal phase and its surroundings. Encapsulating materials have to provide maximum preservation of the active material against environmental conditions, to hold active materials within capsules structure during processing or storage under different conditions, not to react with the encapsulated material, to provide good rheological characteristics at high concentration and simple usage during the encapsulation. In this chapter, encapsulation methods, encapsulation materials, and functions of encapsulation technology in functional beverages were summarized in detail. Biotechnological Progress and Beverage Consumption. https://doi.org/10.1016/B978-0-12-816678-9.00006-0 © 2020 Elsevier Inc. All rights reserved.

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6.2  Encapsulation Methods Encapsulation techniques have been diversified to encapsulate bioactive compounds to increase their bioavailability and stability throughout processing and storage conditions. These methods include traditional methods as spray drying (Đorđević et al., 2015), freeze drying (Ray et al., 2016), extrusion (Prasanna and Charalampopoulos, 2018), coacervation (Comunian et al., 2013), etc. And novel methods as PEGylation (da Silva Freitas and Abrahão-Neto, 2010), nanoemulsions (Silva et al., 2012), and electrospinning (Wen et al., 2017). Spray drying is the most preferred method in food industry systems according to its simple procedure (Drosou et al., 2017). However, there are some disadvantages about efficiency of some mostly used methods. For instance, in the study of López-Rubio et al. (2012), they have observed that encapsulation using spray drying significantly reduced the bacteria viability and stability of bioactive compounds (López-Rubio et al., 2012). Novel techniques were designed in order to rise the efficiency of the encapsulation methods.

6.2.1 Traditional Methods 6.2.1.1  Spray Drying Spray drying is one of the most frequently used encapsulation methods for food industry according to its relatively low cost, reproducibility, and flexibility (Fang and Bhandari, 2010). The disadvantage of this method is the heat applied and accordingly this method cannot be used in bacteria viability and decrease the stability of bioactive compounds compared to novel methods where no heat is applied (Boye, 2015). The procedure of this process involves atomization of homogenized carrier material (1:4) into a drying gas, producing capsules as dry powder that are controlled by product feed, gas flow, and temperature (Kavitake et al., 2017). The schematic diagram is given in Fig. 6.1. In the spray drying process, the payload is preliminarily finely dispersed or homogenized in a highly concentrated (up to 30%wt) biopolymeric aqueous solution, containing starches, succinylated starches, cellulosics, gelatin, gums, and proteins, or in organic solvent solutions, including PLGA, ethyl cellulose, or acrylates, with the main requirement of being able to form a glassy material upon drying. Eventually, another emulsifier might be added to the solution to improve payload dispersion. The dispersion is then sprayed in a drying chamber, forming fine droplets, which are rapidly dried upon contact with a cocurrent or countercurrent flow of hot gas, ultimately forming small micrometric droplets, which are collected in a cyclone or in a filter cloth. The payload of the produced microcapsules is dispersed in the matrix of the encapsulation material (Donsì et al., 2016).

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Fig. 6.1  Schematic illustration of spray drying method (Kavitake et al., 2017).

The basic process of spray drying involves feeding a prepared solution or dispersion of actives into a spray dryer and then atomizing with a nozzle or spinning wheel in a chamber supplied with hot air; the droplet and hot air is contacted in the chamber and the solvent (water) is evaporated from the droplet by the hot air; the dried particles are then separated by a cyclone or bag filter from the humid air and collected in powder form. The spray drying encapsulated particles generally form a matrix structure with a typical spherical shape, and the particle size may vary from very fine (10–50 μm) to large if the spray drying process is integrated with an agglomeration process (Garti and McClements, 2012). The most common use encapsulation materials used in spray drying technique are proteins, carbohydrates, and among others. As a single material may not possess all the functions, mixtures of carbohydrates, gums, and proteins are often used (Garti and McClements, 2012). The encapsulation systems obtained by spray drying method are generally composed of capsules of matrix type, whose properties are mainly affected by the properties of the emulsion and by the process conditions. However, especially for vitamins, aroma compounds, and probiotics, the high temperatures required for the complete solvent evaporation may induce significant thermal damages or volatilization (Boye, 2015). Spray drying is one of the former but also the most preferred application in encapsulation of flavors. The retention of flavors upon

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spray drying also depends greatly on the nature of flavor compounds. In general, the higher retention is associated with the less volatile, the larger molecular weight, and the lower polarity of the flavor compounds (Garti and McClements, 2012). Besides, spray drying method is commonly used in the encapsulation of lipids. Encapsulation of lipids can retard their autooxidation; enhance stability; control lipid-soluble flavor release; mask bitter taste of lipid-soluble substances; and protect dissolved substances against enzyme hydrolysis (Garti and McClements, 2012). Moreover, spray drying has been used for encapsulation of polyphenols to maintain its stability and antioxidant activity. As most of polyphenols are water-soluble compounds, the wall materials used in spray drying encapsulation also need an acceptable level of water solubility, such as starch, maltodextrin (MD), gum arabic, sodium caseinate, or their combinations (Garti and McClements, 2012).

6.2.1.2  Freeze Drying Encapsulation of food ingredients and nutraceuticals by freeze drying is achieved by dissolving, dispersing, or emulsifying these core materials in wall material matrix systems and then co-lyophilizing, usually resulting in a porous, nonshrunken, and uncertain structure. To obtain a product with high quality, some considerations should be kept in mind before the selection of wall materials and preparing the wall/core matrix solutions. The materials should be suitable for their end purpose before being frozen and dried. They should not interact with their own detriment before freezing. Moreover, their desired properties should be sufficiently retained after freezing and freeze drying and the shelf life of the material should be adequate under mild storage conditions (Garti and McClements, 2012). In freeze drying method, most commonly used wall materials are proteins, MDs, disaccharides, gums, and among others. As the freeze-drying process may generate many stresses (e.g., freezing and dehydration) to the cores, the wall materials should have a function to act as a certain degree of cryoprotectant to help stabilize sensitive active agents like probiotics or enzymes. Various special excipients such as buffers (phosphate, tris HCl, citrate, and histidine) and salts (sodium chloride and potassium chloride) are added to protect the system (Garti and McClements, 2012). Freeze drying is performed at low temperatures under vacuum, avoiding water-phase transition and oxidation. The obtained dried mixture must be ground, and the final particles are of large size distribution and with the low surface area. This technology is less frequently used, compared with other encapsulation techniques, as it is very expensive. Addition of cryoprotectants allows reduction of

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cell death during freeze drying of cultures such as probiotic bacterial encapsulates, and stabilizes them during storage. For example, trehalose has been used as a protective coating (Boye, 2015).

6.2.1.3  Fluid Bed Coating Fluid bed coating is based on the deposition of a shell layer on preformed particles such as from spray drying, therefore, it is the final step of production of a core-shell or a coated matrix-type architecture. The particles containing the payload are fluidized, and the coating material is sprayed over them at high pressure, forming a shell layer that is then dried by solvent evaporation or crystallization methods. The coating material usually consists of starches, dextrins, protein derivatives, molasses, lipids, and waxes. Prior to spraying, the coating material is either melted or dispersed in a suitable solvent that can be easily evaporated, forming a viscous system with an enhanced tendency to deposition and adhesion on the payload particles. The air or gas flow through the fluidized bed serves to chill the molten material, or to evaporate the solvent, causing its consolidation in a shell layer (Donsì et al., 2016).

6.2.1.4 Emulsions Emulsion encapsulation involves dispersing the compound of interest including essential oils, flavors, omega-3 fatty acids, and antioxidants in an aqueous solution of a “film-forming” polymer, usually a carbohydrate, which, upon drying (usually spray drying), would produce a polymer matrix containing the bioactive component. Encapsulant wall materials for this purpose include gum arabic, MD, modified starches, and celluloses (Boye, 2015).

6.2.1.5  Spray Chilling Spray chilling, represents an alternative to spray drying, used to prevent the volatilization or degradation of thermolabile food additives. Spray chilling is also based on the preliminary dispersion of the payload in a solution, that can be made of low molecular weight polymers, resins, hydrogenated vegetable oils or waxes, and its atomization, with the capsule consolidation not being based on dehydration, but on glass transition or crystallization of the encapsulant material upon rapid chilling in a cooled gas flow (Donsì et al., 2016). In spray chilling, a molten matrix with low melting point containing the bioactive substance is atomized through a pneumatic nozzle into a vessel. This technology is, in principle, opposite to spray drying instead of evaporating, the dispersion containing the bioactive material is cooled to allow immobilization. Cold air is injected into the

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vessel to enable solidification of the gel particle. The liquid droplet solidifies and entraps the bioactive substance. This technology is rarely used to microencapsulate live probiotic organisms, however, it is more suitable for encapsulation of vitamins, fatty acids, antioxidants, yeasts, and enzymes (Boye, 2015).

6.2.1.6 Extrusion The extrusion process is conducted using a biopolymer solution including matrix materials forced through a nozzle into a gelling solution at low temperature. In laboratory conditions, extrusion is performed in a syringe, where the biopolymer solution is loaded and extruded through the needle (Joye and McClements, 2014). This equipment can be modified using direct current (Li et al., 2016). The most commonly used extrusion system includes sodium alginate (SA) into a calcium chloride solution (Munin and Edwards-Lévy, 2011). The advantages of this method are processing at mild conditions, wide range of usage, and stability of gel particles through storage (Gouin, 2004). The disadvantages of this method are producing large and porous particles enabling encapsulated particles to diffuse, few options of matrix materials, difficult, and high-cost process (Gouin, 2004).

6.2.1.7 Coextrusion Coextrusion technology is instead used to produce core-shell particles and is based on the extrusion through a concentric nozzle, with the payload dispersion being extruded through the inner nozzle and the wall materials being extruded from the outer nozzle. Owing to its ability to fabricate in a simple and robust process encapsulation systems with multiple coating layers (using extrusion nozzles with multiple concentricities), coextrusion is used when a slow and controlled release of the payload, as well as taste masking is desired. Particle consolidation occurs either through chilling and glass transition of the wall material, gelation, or evaporation of the solvent. In the first case, the same coating materials of melt extrusion are used, while in the other cases, viscous polymer solutions are used, based on proteins and polysaccharides, gums, and other commercial polymers. The main disadvantages of melt extrusion and coextrusion processes are the high temperatures and the high shear rates attained in the extruder (Donsì et al., 2016). The coextrusion process is different from classical extrusion, which is a separate technique often used for encapsulation. The classic extrusion process typically refers to the use of pressure and elevated temperature to force a molten emulsion or solution through a die and subsequently cooled to form a fiber or rod, which is then ground into

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a fine powder. Extrusion is used for the encapsulation of flavors to produce a matrix particle. Coextrusion requires the use of liquids that may be room temperature and is extruded out of a concentric nozzle in the absence of high pressures (Garti and McClements, 2012).

6.2.1.8 Coacervation Coacervation is a widely used encapsulation method to produce capsules as shown in Fig. 6.2. First of all, one or a mixture of polyelectrolytes were separated in a solution to form coacervate phase. Afterward, this coacervate phase sheeted around the compound to be encapsulated (Ezhilarasi et al., 2013). If it is formed by one type of polymer it is called simple coacervation, whereas if there is more than one type of polymer it is named as complex coacervation. The produced shell can be cross-linked by a chemical or enzymatic cross-linker to enhance its robustness (Ezhilarasi et  al., 2013). The propulsive force is strong electrostatic interactions in coacervate production, but they should not be extra strong as to prevent precipitation of coacervates (Augustin and Hemar, 2009; Munin and Edwards-Lévy, 2011). Besides, hydrophobic interactions and hydrogen bonds also carry importance in coacervation (Joye and McClements, 2014). The advantages of the coacervation method are that it has a high encapsulation efficiency (Ezhilarasi et  al., 2013). However, there are various disadvantages of this method including instability in aqueous solutions (Joye and McClements, 2014), higher cost (Munin and Edwards-Lévy, 2011), and sensitivity to pH (Augustin and Hemar, 2009).

6.2.1.9 Liposomes Liposomes are often employed as delivery systems of bioactive compounds, as during the formation of the liposome, hydrophobic material may be incorporated in the lipid membrane while hydrophilic molecules present in the aqueous phase may become trapped inside the liposome. Thus, liposomes can encapsulate both hydrophilic and

Fig. 6.2  Production of a capsule by coacervation method.

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hydrophobic compounds within a single structure. Although these phospholipid vesicles show great potential for encapsulation and controlled release of nutritionally significant compounds, their application in foods has yet to be fully exploited. Liposomes protect their contents from the external environment while still allowing small ­molecules to permeate in and out of the membrane. They imitate living cells, and have been used to model the structure and function of biological membranes, and to study drug intracellular uptake. They are biologically compatible and have been used in a broad range of pharmaceutical and cosmetic systems. Liposomes can also be employed to control or delay the release of the encapsulated material (Garti and McClements, 2012). Liposomes are produced by the association of amphiphilic compounds (mainly phospholipids) into bilayer structures. A liposome is defined as a structure composed of lipid bilayers that enclose a number of aqueous or liquid compartments. Using various techniques such as solvent evaporation, ultrasonication, and microfluidization, the bilayer forms spherical core-shell structures (vesicles). During the formation process, hydrophilic molecules in the external aqueous phase become entrapped in the liquid regions and core, while hydrophobic material may be incorporated in the bilayer membranes. The release of the entrapped substance can be either a gradual process resulting from diffusion through the membranes, oral most instantaneous following membrane disruption caused by changes in pH or temperature. Liposomes are mainly used for delivery of lipids or water-soluble materials such as omega-3 fatty acids, yeasts, and enzymes (Boye, 2015).

6.2.2 New Methods 6.2.2.1 PEGylation The covalent bonding of polyethylene glycol (PEG) to a given molecule is named as PEGylation It is a novel method mostly used in the field of targeted delivery of bioactive compounds. The process of PEGylation involves liposomes, peptides, carbohydrates, antibody fragments, nucleotides, small organic molecules, and different nanoparticle (NP) formations (Mishra et al., 2016).

6.2.2.2 Nanoemulsions Nanoemulsions can be explained as thermodynamically unstable colloidal dispersion including two immiscible liquids and one liquid should be dispersed as small spherical droplets with mean radii between 100 nm and 100 mm (McClements and Rao, 2011). This method is more advantageous compared to traditional emulsion method

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a­ ccording to their smaller droplet size. These advantages can be listed as: (i) high optical clarity, (ii) high physical stability toward gravitational separation and droplet aggregation, and (iii) improved bioavailability of encapsulated materials, which make them appropriate for food processes (Montes de Oca-Ávalos et al., 2017). This method carries an importance in the functional beverage industry as they have small droplet sizes that ensure a weak light scattering of final products and therefore, the addition of bioactive compounds to functional food beverages will not cause any problems for clarification (Wang et al., 2016). The physicochemical and structural properties of NPs should be controlled to achieve a desired beverage application in delivery of nutraceuticals, vitamins, drugs, antimicrobials, colors, or flavors (Esmaeili and Gholami, 2015; Mehmood, 2015).

6.2.2.3 Electrospinning Electrospinning technique has been used to encapsulate bioactive compounds according to the advantages of this method compared to traditional techniques. The most important advantages of this method are that there is no heat application and therefore, the structures of the bioactive compounds are preserved. It is also an easy and flexible method to produce fibers with high surface-to-volume ratio and porosity and therefore, the efficiency of encapsulation is enhanced. Electrospinning method is applied using an electrical field to continuously draw the droplet of polymer solution or melt polymer into a fine fiber followed by its deposition on a grounded collector (Wen et al., 2017). A schematic illustration of electrospinning system is shown in Fig. 6.3.

Fig. 6.3  Schematic illustration of the basic setup for electrospinning (Nieuwland et al., 2013).

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6.3  Encapsulation Materials A large variety of materials can be used as encapsulation of food components including MDs, gum arabic, hydrophobically modified starches and chitosan, as well as mixtures of them, the most commonly used coating materials.

6.3.1 Maltodextrins MDs are widely used as encapsulation material in functional beverages according to their low cost, neutral taste and aroma, and also their role of protection against flavors (Fernandes et al., 2014).

6.3.2 Gum Arabic Gum arabic is another polysaccharide alternative to MDs, widely used in encapsulation processes in functional beverages due to its good emulsifying and film-forming characteristics (Silva et al., 2013).

6.3.3  Hydrolyzed Starches This is one of the most common wall or carrier materials. The hydrolyzed starches are available in dextrose equivalent (DE) ranging from 2 to 36.5 and offer good protection against oxidation. These are low in viscosity at high total solid contents. However, they lack in emulsifying properties. It is, therefore, used along with gum acacia or other emulsifying agents like protein, whey protein isolates, and whey protein concentrates. MD and low DE corn syrup solids (CSSs) when dried show matrix forming properties important in the wall system. When MDs or CSS are used as wall constituents, it is necessary to incorporate other wall material such as gelating agent, sodium caseinate, whey proteins, lecithins, etc. for improving emulsifying characteristics (Kuna and Poshadri, 2010).

6.3.4 Chitosan Chitosan and its derivatives are another types of polysaccharides that are used commonly in functional beverages in order to encapsulate bioactive compounds. Chitosan is used widely in functional beverage applications as a coating material of bioactive compounds for preventing oxidation and improving the bioavailability of probiotics in gastrointestinal system. Chitosan coatings are used in functional beverages to protect bioactive compounds including terpenes, carotenoids, anthocyanins (He et  al., 2017; Djordjevic et  al., 2007), and probiotics (Nualkaekul et  al., 2012; Krasaekoopt et  al., 2006) from acidic conditions of GIT and also environmental conditions.

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They also enhance the solubility of lipophilic compounds as carotenoids (García-Márquez et al., 2015) and mask unacceptable taste to improve products’ acceptability by consumers (Souza et al., 2014). Nualkaekul et al. (2012) have been used chitosan-coated alginate beads for the protection of microencapsulated Lactobacillus planta­ rum in pomegranate juice. He et  al. (2017) also used anthocyanin-­ loaded chitosan NPs in a model beverage system and observed that compared with the free anthocyanin solutions in a model beverage system, the stability of the anthocyanins was increased in the anthocyanin-loaded chitosan NPs.

6.3.5 Calcium or SAs Calcium or SAs are polysaccharides derived from algae, which are frequently used to encapsulate probiotics, because of their lack of toxicity, biocompatibility, low cost, and ease of use. Upon drying, they form a porous structure, which is not resistant to acidic environments, and therefore, is not suitable to provide sufficient stability in the gastric tract. However, enhanced stability can be achieved by blending calcium alginate with other biopolymers, or further coating alginate capsules with a layer of insoluble polymers (Donsì et al., 2016).

6.3.6 Gellan and Xanthan Gums Gellan or xanthan gums are polysaccharides of microbial origin, obtained from Pseudomonas elodea and Xanthomonas campestris, respectively. Their mixture was reported to be extremely suitable to encapsulate probiotics in systems with high resistance in acidic environments (Donsì et al., 2016).

6.3.7  K-Carrageenan K-carrageenan is a natural polymer, widely used in the food industry, which is extremely compatible with microbial cells, ensuring high viability after the encapsulation process. However, the resulting gel structures have limited physical stability, upon the stress conditions experimented in food transformation, requiring its blending with other polymers (Donsì et al., 2016).

6.3.8  Whey Proteins As starch and related products lack emulsification properties, they are used as wall materials along with surface-active wall constituents. Whey protein owing to their structure gives functional properties desired for effective microencapsulation of an hydrous milk fat. Whey protein in combination with MDs and CSSs is reported to be the

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most effective encapsulation material during spray drying (Kuna and Poshadri, 2010).

6.3.9  Mixed Coatings Mixed coatings have been used to encapsulate bioactive compounds like essential oils (Fernandes et  al., 2014), anthocyanins (Flores et  al., 2014; Mahdavi et  al., 2014), cherry pomace phenolic ­extracts (Cilek et al., 2012), and among others.

6.4  Functions of Encapsulation Technology in Functional Beverages 6.4.1 Aroma and Flavor Stabilization Aroma and flavors are important ingredients in functional foods, but they face some problems of instability, losses during heating and reactions with other matrix ingredients in the beverage industry. They have altered these problems by using encapsulation technology. Cyclodextrins provide a protective barrier to aroma and flavor compounds that reduce their instability, losses during heating and reactions with other matrix ingredients (Marques, 2010).

6.4.2 Increasing Bioavailability and Targeted Delivery of Phenolic Compounds Bioavailability depends on various factors such as food processing, with the host organism (sex, age, and composition of intestinal microflora), as well as on interactions between polyphenols and other molecules such as salivary proteins and digestive enzymes. It is essential to better known bioavailability of the phenolic compounds of functional beverages to establish evidence for the effects of consumption on human health and to better identify which polyphenols provide the greatest effectiveness. Encapsulation is an effective technique to increase the bioavailability and result in targeted delivery of phenolic compounds. The details on the bioavailability of encapsulated-bioactive compounds in functional beverages are very limited. Tomás-Navarro et  al. (2014) encapsulated hesperidin which is a flavanone abundant in orange juice using gum arabic as a coating material together with micronization technique. They have found that micronization (5.1 μm) increased flavanone’s bioavailability twofold compared to conventional hesperidin (32.8 μm) (Tomás-Navarro et al., 2014).

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Moreover, Motilva et  al. (2016) have studied the impact of nano-­ encapsulation on wine polyphenols. A dealcoholized red wine was used as the vehicle for enrichment with both nonencapsulated and nano-encapsulated phenol extracts. They observed that nano-­ encapsulation of the extract slightly enhanced the bioavailability of malvidin-3-O-glucoside, as observed in the higher urine excretion of its native form and its microbial metabolites syringic and gallic acids (Motilva et al., 2016). Furthermore, Li et al. (2017) investigated the encapsulation of green tea catechin derivative, lycopene, in PEG NPs coated with c­ hitosan. Lycopene-loaded NPs were prepared by nanoprecipitation followed by coating within chitosan to form a shell. Chitosan was coated onto the surface of lycopene NPs because chitosan exhibits pH-dependent behavior which allows overcoming the harsh environment. To understand the pH-dependent behavior effect on the release kinetics of lycopene, time-dependent release of lycopene in simulated gastric fluid and simulated intestinal fluid was studied. A known amount of lyophilized NPs were dispersed in simulated gastric juice or simulated intestinal fluid. In simulated gastric fluid, NPs released 5% of the total lycopene compared to 12% of total lycopene released in the simulated intestinal fluid in 24 h. In all, 7% of total lycopene was released in the burst-release phase from NPs in simulated gastric fluid compared to 16% of total lycopene released in the simulated intestinal fluid in 24 h, both formulations showed pH-dependent release. Authors proposed that chitosan release is based on lysozyme degradation and swelling (Li et al., 2017). Besides, Ruiz-Rico et al. (2017) examined the bioaccessibility of the folic acid during the simulated digestion of orange and apple juices, using inorganic encapsulation systems, for example, mesoporous silica particles (MSPs). They have observed that folic acid was preserved in the salivary and gastric period, and vitamin bioaccessibility was improved in the intestinal period (Ruiz-Rico et al., 2017).

6.4.3 Increased Antioxidant Activity Zokti et al. (2016) studied the effect of green tea catechin extracts microparticles produced by spray drying method with different Wall materials including MDs, gum arabic, and chitosan on antioxidant activity of mango drinks. They have observed that the encapsulated catechins compounds were more stable in the supplemented mango drinks in comparison with the non-encapsulated catechin powder with improved functionality. They have also observed that antioxidant activity of the mango drinks during storage was proportional to the increase in the concentration of microparticles incorporated (0.5%, 1.0%, and 2.0% w/v) (Zokti et al., 2016).

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Ballesteros et al. (2017) used coating materials including MD, gum arabic, and a mixture of these components for the encapsulation of antioxidant phenolic compounds extracted from spent coffee grounds using both freeze drying and spray drying methods. They have observed that 73%–86% of the antioxidant activity present in the original extract was preserved by encapsulation of phenolic compounds (Ballesteros et al., 2017).

6.4.4 Shelf Life Extension One of the major drawbacks, which limit the application of bioactive compounds in functional beverages, is their law stability during storage. The applications of encapsulation techniques can provide a solution to this problem and increase the shelf life of the functional food beverages. Naghavi et al. (2016) have studied the effect of green tea extracts nanoliposomes to expand the shelf life of fresh orange juice and pomegranate juice. Green tea extract nanoliposomes were produced by ethanol injection method. The juices quality was determined by their ascorbic acid content through 30 days of storage period. A significant decrease in ascorbic acid was observed in control samples with no green tea nanoliposomes were added. It was concluded that the antioxidant activity exhibited by the green tea nanoliposomes was responsible for the extended shelf life of the juices. Therefore, incorporating green tea extract nanoliposomes could eliminate the destruction of vitamin C upon long storage time in fruit juices (Naghavi et al., 2016). Moreover, Sawale et al. (2017) studied the effect of the addition of free and encapsulated forms of Terminalia arjuna on storage stability of chocolate vanilla dairy drink. They have observed that the physicochemical changes occurred at a slower rate in chocolate vanilla dairy drink that contains encapsulated form of T. arjuna herb. Viscosity and sedimentation stability were significantly (P < .05) higher than the control sample during the entire storage period. It was recommended that supplementation of T. arjuna herb in encapsulated form improve the stability of dairy drinks during storage period (Sawale et al., 2017).

6.4.5 Protection of Valuable Nutritious Compounds 6.4.5.1  Unsaturated Lipids Functional beverages may be enriched with encapsulated unsaturated lipids according to their beneficial health effects. Ilyasoglu and El (2014) added fish oil as a source of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) into fruit juices within a multilayered interfacial membranes using electrostatic attraction between sodium caseinate and gum complexes. After an in vitro digestion (EPA, 56.16%;

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DHA, 36.25%; and eicosapentaenoic + DHAs, 47.37%), the bioaccessibility of this system was found to be good. Accordingly, NP fortification into fruit juice did not change the sensory quality of functional fruit juices. Furthermore, Habibi et al. (2017) investigated the adaptability of microcapsules, prepared by gelatin-gum arabic coacervates, such as fish oil within pomegranate juice. Fish oil was regularly released from beads (14% after 42 days). However, they have found that the addition of microcapsule powder to pomegranate juice resulted in a negative impact on the sensory acceptability opposite of the study of Ilyasoglu and El (2014).

6.4.5.2  Phenolic Compounds Phenolic compounds are secondary metabolites found abundantly in a wide variety of foods, such as fruits, vegetables, herbs, seeds, and cereals, and in beverages, such as coffee, tea, fruit juices, cocoa, and wine (Vinson et al., 2001). They are currently a topic of great scientific attention due to the interest in their potential health benefits, which include anticancer, antioxidant, antimicrobial, and antiinflammatory properties. Polyphenols are also implicated in preventing chronic diseases such as cardiovascular diseases, diabetes, obesity, and neurodegenerative diseases, among others (Scalbert et al., 2005; Khurana et al., 2013; Meydani and Hasan, 2010; Li et al., 2014). However, they are very vulnerable to oxidation as they have unsaturated bonds in their structure according to conditions as light, oxygen, and moisture, among others. Therefore, encapsulation of these valuable compounds carries great importance to increase their storage stability as well as producing functional foods that have health-promoting effects to consumers. Besides, there is another reason of encapsulation of phenolic compounds as masking unpleasant flavors of these compounds as many of them provide a bitter taste and astringency to food products (Ray et al., 2016). Table 6.1 gives a summary of recent usage of encapsulation technologies on phenolic compounds used in functional beverage products. Ballesteros et al. (2017) used coating materials including MD, gum arabic, and a mixture of these components for the encapsulation of antioxidant phenolic compounds obtained from spent coffee grounds using both freeze drying and spray drying methods. As a result, they have found that freeze drying using MD as the coating material is the optimum encapsulation method for antioxidant phenolic compounds extracted from spent coffee grounds. Under these conditions, 62% and 73% of phenolic compounds and flavonoids were retained in encapsulated spent coffee ground samples, respectively (Ballesteros et  al., 2017). Aditya et  al. (2015) encapsulated catechin and curcumin to pre­ vent their degradation in beverage systems. The stability of catechin was increased by >20% at 23 ± 2°C and by >40% at 4°C after 15 days of

Table 6.1  Encapsulation of Phenolic Compounds in Functional Beverages Functional Beverage Product

Encapsulated Phenolic Compounds

Spent coffee grounds

Phenolic compounds extracted from coffee

Model beverage system

Catechin curcumin

Model beverage system

Encapsulation Material

Encapsulation Technique

Maltodextrin, gum arabic, and a mixture of these components (ratio 1:1) Olive oil, soybean oil, and sunflower oil

Blueberry-derived mixture of anthocyanins

Carboxymethyl chitosan

Hibiscus extract

Anthocyanins

Model beverage systems

Catechin

Emulsion (w/o) was produced at first with rapeseed oil and polyglycerol polyricinoleate (PGPR) surfactant β-cyclodextrin

Produce nanoparticles through ionic gelation Ionic gelation (IG) using two techniques: dripping-extrusion and atomization

Function

References

Freeze-drying Spray-drying

Preserving phenolic components providing the highest retention percentages of phenolic compounds and flavonoids within the matrix

Ballesteros et al., 2017

Water- in-oil (W/O) emulsion

The stability of catechin was increased by >20% at 23 ± 2°C and by >40% at 4°C after 15 days of incubation, compared to free catechin. Curcumin stability was increased by >80% in the beverage system in the form of emulsions, whereas it was reduced to ~40% in the case of free curcumin. Protection of anthocyanins against degradation under thermal conditions

Aditya et al., 2015

Increased thermal stability of anthocyanins

de Moura et al., 2018

Increased stability of catechin

Ho et al., 2017

Complexation

He et al., 2017

Bayberry juice

Anthocyanins

Whey protein isolate and maltodextrin

Spray-drying

Increased stability of anthocyanins

Pomegranate juice Sour cherry juice

Anthocyanins

Soybean protein isolate and maltodextrin Maltodextrin gum arabic

Spray-drying

Increased stability of anthocyanins

Spray-drying

High-performance agave fructans (HPAF) and high degree of polymerization agave fructans (HDPAF) and maltodextrin (MD) –

Spray-drying

Maltodextrin is optimal wall material for encapsulation of sour cherry juice at temperature of 200°°C, Flavonol glycosides were retained to the most (84.01%) in juice encapsulated at 180°C with gum arabic added in ratio 2:1 The highest yield and concentration of anthocyanins after drying and during storage were found for a 1:6 core:wall material ratio

Phenolic acids, anthocyanins and flavonol glycosides

Pitanga (Eugenia uniflora L.) juice

Apple juice

Steppogenin

Apple juice Orange juice Apple juice

Folic acid

Apple juice

Pentapeptide (rice bran derived) Chlorogenic acid

Grape juice

Artocarpanone and ascorbic acid

Mesoporous silica particles ethyl butyrate, Ethyl oleate, dl-α-tocopherol, soybean oil, isopropyl myristate, caprylic capric triglyceride Poly (lactic-co-glycolic acid) β-Cyclodextrin and 2-hydroxypropyl)-βcyclodextrin

Oil-in-water microemulsion Impregnation

High inhibition of juice browning

Oil-in-water microemulsion

High inhibition of juice browning

Nano-precipitation

Increased stability for 2 months of storage time Decrease in degradation of anthocyanins

Inclusion complexation

Increased stability and controlled release

Fang and Bhandari, 2011 Robert et al., 2010 Elez Garofulić et al., 2017

Ortiz-Basurto et al., 2017

Tao et al., 2017 Ruiz-Rico et al., 2017 Dong et al., 2016

Alessa et al., 2014 Shao et al., 2014

Continued

Table 6.1  Encapsulation of Phenolic Compounds in Functional Beverages—cont’d Functional Beverage Product

Encapsulated Phenolic Compounds

Orange juice Skim milk

Astaxanthin

Orange nectar

(−)-epigallocatechin gallate

Encapsulation Material

Encapsulation Technique

Nanodispersions prepared using sodium caseinate, polysorbate, gum Arabic or optimum formulated three component stabilizer system Liposomes or alginate or chitosan microparticles reinforced with liposomes

Function

References

Emulsificationdiffusion

Increased bioavailability of astaxanthin

Anarjan and Tan, 2013

Liposome

Decrease rate of degradation of (−)-epigallocatechin gallate

Istenič et al., 2016

Chapter 6  Role of Encapsulation in Functional Beverages   213

incubation, compared to free catechin. Besides, cucumin stability was increased by >80% in the beverage system in the form of emulsions, whereas it was reduced to ~40% in the case of free curcumin (Aditya et al., 2015). Ho et al. (2017) studied encapsulation of catechin in model beverage systems. Catechin has an astringent taste, less solubility in water, and susceptible to oxygen, light, and alkaline conditions which limit its application in functional food beverages. They have observed that stability of catechin was significantly enhanced by β-cyclodextrin encapsulation and suggested to be used in functional beverage production (Ho et al., 2017). For stabilization of anthocyanins, Fang and Bhandari (2011) spraydried bayberry juice using whey protein isolate and MD matrices. Moreover, Robert et  al. (2010) spray-dried pomegranate juice to increase the stability of anthocyanins using soybean protein isolate and MD matrices. Recently, de Moura et  al. (2018) microencapsulated Hibiscus extract anthocyanins by using ionic gelation method. They have observed that encapsulation of anthocyanins using ionic gelation resulted in higher temperature stability compared with the free extract (de Moura et al., 2018). Recently, Elez Garofulić et al. (2017) studied the retention of phenolic acids, anthocyanins, and flavonol glycosides in sour cherry Marasca juice encapsulated by spray drying as affected by temperature, type of wall material, and its ratio to the juice dry matter. They have observed the amount of individual polyphenols using high-­ performance liquid chromatography (HPLC) and response surface methodology approach in order to find the differences between different groups of sour cherry polyphenols concerning the conditions performed for encapsulation process. They have observed that the optimal wall material for encapsulation of sour cherry juice at temperature of 200°C through the use MD as a wall material in ratio 3:1, with the highest retention of phenolic acids and anthocyanins, 93.31% and 88.68%, respectively. Adversely, flavonol glycosides were retained to the most (84.01%) in juice encapsulated at 180°C with gum arabic added in ratio 2:1 (Elez Garofulić et al., 2017). Another recent study to improve stability of anthocyanins was performed by Ortiz-Basurto et al. (2017) who microencapsulated pitanga (Eugenia uniflora L.) juice by spray drying, using high-­performance agave fructans (HPAF) and high degree of polymerization agave fructans (HDPAF), and MD, respectively, as the wall materials. They have evaluated the antioxidant and physicochemical properties of the capsules during storage at various temperatures. The microparticles developed using fructans HPAF and HDPAF, exhibited similar physicochemical and flow properties to those presented by the microparticles prepared with MD. The highest yield and concentration

214  Chapter 6  Role of Encapsulation in Functional Beverages

of anthocyanins after drying and during storage were found for a 1:6 core: wall material ratio (Ortiz-Basurto et al., 2017). Moreover, Tao et al. (2017) prepared oil in water microemulsions of a natural flavanone steppogenin that has high tyrosinase inhibitory capacity to decrease browning of fresh apple juice. They have found that steppogenin microemulsion stop browning of apple juice for 24 h at room temperature and 7 days at 4°C (Tao et al., 2017). There are some recent studies on effect of encapsulating bioactive compounds in apple juice (Tao et al., 2017; Ruiz-Rico et al., 2017; Alessa et  al., 2014; Dong et  al., 2016). As a result of these studies, it can be stated that encapsulating bioactive compounds can decrease browning in apple juice, improve the stability of bioactive compounds. Moreover, Shao et al. (2014) encapsulated chlorogenic acid using inclusion complexation method and observed reduced degradation of anthocyanins. Anarjan and Tan (2013) used orange juice, skimmed milk as pattern food systems, and in deionized water as a control to study the stability of nanodispersions of astaxanthin. Astaxanthin was less bioavailable from diluted nanodispersions in juice compared with those diluted in deionized water, due to the acidic pH of orange juice. Istenič et al. (2016) observed the stability of (−) epigallocatechin gallate (EGCG) encapsulated in liposomes and the chitosan and alginate microparticles entrapped with liposomes within orange nectar. The encapsulated EGCG was more stable as 6% of the entrapped core was degraded after 14 days of storage time, whereas the control EGCG was degraded about 30% or more.

6.4.5.3 Vitamins There is a growing demand of consumers in food products that contain bio-functional ingredients that are beneficial for human health. It is desirable to add vitamins into food systems as a functional ingredient, but it is also challenging for several reasons. As an example, the incorporation of vitamin E in functional beverages is difficult, because it is a highly lipophilic compound that cannot be dispersed into aqueous-based food matrices (Sagalowicz and Leser, 2010). It is also prone to oxidation and can be degraded through processing and storage (Gawrysiak-Witulska et al., 2009; Yoon and Choe, 2009). Vitamin E can be incorporated into a colloidal dispersion consisting of small lipophilic particles to disperse in aqueous solutions (Piorkowski and McClements, 2014). Table  6.2 gives a summary of recent applications of encapsulation technologies on vitamins used in functional beverage products. Recently, Raikos (2017) has studied the effects of thermal processing on the stability of orange oil beverage emulsion containing vitamin E during chilled storage. It was observed that vitamin E retention was

Table 6.2  Encapsulation of Vitamins in Functional Beverages Functional Beverage Product

Encapsulated Phenolic Compounds

Orange oil beverage emulsion

Vitamin E

Model beverage system

Panthotenic acid

Apple juice

Ascorbic acid

Orange juice

Orange juice

Encapsulation Material

Encapsulation Technique

86.78% water, 3.0% WPI, 3.5% orange oil, 3.0% sweetener, 2.0% fiber, 0.7% citric acid, 1.0% coloring agents, 0.02% tocopherols Alginate or alginate epectin mixtures

Emulsion

Liposomes

Vitamin E and vitamin C

Dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylcholinecholesterol Soy-phosphatidylcholine-based liposomes

Vitamin E and vitamin C

Soy-phosphatidylcholine-based liposomes

Dehydration-rehydration

Lyposomes/hydrogel microcapsule

Dehydration-rehydration

Function

References

Vitamin E retention was considerably higher for all heated beverages (85%) under the specified storage conditions Encapsulated panthothenic acid showed great stability, whereas they also resulted in greater retention at higher temperatures, compared to liposomes kept at lower pH values Decreased oxidation rate of ascorbic acid

Raikos, 2017

Liposomes show great stability and protection of vitamin C Liposomes show great protection on antioxidan activity of vitamin C and vitamin E before and after pasteurization

Marsanasco et al., 2015

Ota et al., 2018

Wechtersbach et al., 2012

Marsanasco et al., 2011

216  Chapter 6  Role of Encapsulation in Functional Beverages

considerably higher for all heated beverages (85%) under the specified storage conditions (Raikos, 2017). Recently, Ota et al. (2018) encapsulated pantothenic acid in liposomes and in various hydrogel microcapsules that consisted of alginate or alginate pectin mixtures to increase its stability. They have reached encapsulation efficiency of 0.75 ± 0.02 using liposomes and 0.60 ± 0.02 using alginate microparticles. At moderate pH (4.0) conditions, liposomes with encapsulated pantothenic acid showed great stability, whereas they also resulted in greater retention at higher temperatures, compared to liposomes kept at lower pH values (Ota et al., 2018). Moreover, Wechtersbach et al. (2012) encapsulated ascorbic acid using liposome method and observed that the rate of degradation of encapsulated ascorbic acid in apple juice at 25°C was reduced by almost twofold when entrapped in liposomes. Marsanasco et al. (2011) encapsulated vitamins E and C using dehydration-rehydration method with soy-phosphatidylcholine-based liposomes in orange juice. They have observed high protection in antioxidant capacity of vitamins E and C. In their further study, they have observed higher stability of and important protective effect on ascorbic acid (Marsanasco et al., 2015).

6.4.5.4 Probiotics Probiotic cells were encapsulated to increase their viability protecting them from the acidic pH of various functional beverage products. Encapsulation methods have been widely researched to create a physical barrier protecting the bacteria from adverse conditions during production processes and digestion (Fritzen-Freire et al., 2012). Table 6.3 gives a summary of recent applications of encapsulation technologies on probiotic microorganisms used in functional beverage products. Encapsulation using alginate increased the cell viability (Anekella and Orsat, 2013; Ding and Shah, 2008). The protection of probiotic cells (Nualkaekul et  al., 2012) was increased by chitosan-coated alginate beads. Krasaekoopt and Watcharapoka (2014) investigated the cell viability of the probiotic cultures Lactobacillus casei and Lactobacillus ac­ idophilus and concluded that the cell viability of the probiotic cultures Lactobacillus casei and Lactobacillus acidophilus has grown with the inclusion of galactooligosaccharides into the encapsulating matrices. Chaikham (2015) examined the effect of alginate encapsulation with Thai herbal extracts including yanang, cashew flower, and pennywort on viability of probiotic bacteria suspended in fruit juices such as mulberry, maoberry, longan, and melon juices. They have observed that green tea and cashew flower extracts noticeably enhanced the stability of probiotic beads in all the products, as compared to the controls during storage (Chaikham, 2015). Besides Chaikham et al. (2013) used emulsification method to encapsulate Lactobacillus acidophilus. They

Table 6.3  Encapsulation of Probiotics in Functional Beverages Functional Beverage Product

Encapsulated Microorganisms

Encapsulation Material

Encapsulation Technique

Function

References

L. casei 01, L. acidophilus LA5 and B. lactis Bb-12

Alginate-Thai herbal extracts

Extrusion

Enhanced stability of probiotic beads

Chaikham, 2015

L. acidophilus LA5

Alginate

Emulsion

Chaikham et al., 2013

Green tea extract Goat milk Cow milk

Lactobacillus helveticus Bifidobacterium longum subsp. infantis CCUG 52486

Whey protein-coated pectinate microparticles Sodium alginate (SA), sodium alginate-cow milk (SACM), sodium alginategoat milk (SAGM) and sodium alginate-casein hydrolysate (SACH)

Co-encapsulation

Increased viability of L. acidophilus LA5, increase in short-chain fatty acids concentration (acetate, propionate, and butyrate) Improved viability of probiotic bacteria

Apple juice

L. rhamnosus GG

Whey protein isolate (WPI), mixture with a physically modified starch (WPI-RS) or resistant starch (RS)

Longan juice Maoberry juice Melon juice Mulberry juice Wild cherry Longan juice

Extrusion

Spray drying

SACM and SAGM are suitable to encapsulate B. longum subsp. infantis CCUG 52486 using the extrusion technique SAGM has a potential to be used as a new encapsulation material for encapsulating probiotic bacteria, resulting milk and goat milk-based products with higher probiotic cell concentrations during refrigerated storage The bacteria are isolated from the stresses of the low pH of apple juice over 5 weeks of storage at 25°C

Gaudreau et al., 2016 Prasanna and Charalampopoulos, 2018

Ying et al., 2013

Continued

Table 6.3  Encapsulation of Probiotics in Functional Beverages —cont’d Functional Beverage Product

Encapsulated Microorganisms

Encapsulation Material

Encapsulation Technique

Acerola nectar

B. animalis subsp. lactis BB-12

Cellulose acetate phytalate

Spray drying

Apple juice

L. rhamnosus GG

Extrusion

Carrot juice

L. casei 01

Cowberry juice

S. cerevisiae var. boulardii Lactobacillus plantarium (NCIMB 8826) and Bifidobacterium longum (NCIMB 8809) L. rhamnosus GG

Chitosan-alginate with/ without inulin Chitosan-Ca-alginate enriched with fructooligosaccharide Alginate-inulin xanthan gum Alginate or pectin with a chitosan, gelatin or glucomannan

Cranberry juice

Cranberry juice Pomegranate juice

Grape juice

L. casei 01

Grape juice Orange juice Pineapple juice

B. adolescentis (ATCC 15703)

Whey protein coated with apple, citrus pectin, alginate, k-carrageenan, iota-carrageenan and inulin Alginate with chitosan coating Pea protein-alginate mixture

Spray-drying

Function

References

Spray-dried bacteria count was higher (8 log CFU/mL) after 30 days of storage compared to control (5.9 log CFU/mL) 4.5 times higher viability of L. rhamnosus GG compared to control Enhanced viability of L. casei 01 in carrot juice

Antunes et al., 2013 Gandomi et al., 2016 Petreska-Ivanovska et al., 2014

Extrusion

Enhanced cell viability

Extrusion

Enhanced cell viability (highest was reached using double gelatin coated pectin beads)

Extrusion

Enhanced cell viability (8.6 log CFU/mL) after 28 days of storage

Doherty et al., 2012

Extrusion

Consumer acceptance and enhanced cell viability Improved cell viability during storage at 22°C

Krasaekoopt and Kitsawad, 2010 Wang et al., 2015

Water-in-oil emulsion

Fratianni et al., 2014 Nualkaekul et al., 2013

Alginate, with chitosan or dextran sulfate or without double coatings Alginate

Extrusion

Improved cell viability (9 log CFU/mL) after 50 days of storage time at 5°C

Rodrigues et al., 2012

Impinging

Reduced acidification and negative sensory effect of probiotics

Sohail et al., 2012

Alginate with chitosan coating

Extrusion

Improved cell viability (8 log CFU/mL) after 4 weeks of storage time at 4°C

L. casei 431 and L. acidophilus La-5 B. longum (NCIMB 8809) and B. breve (NCIMB 8807)

Alginate

Extrusion

Bacterial poly-γ-glutamic acid

Freeze drying

L. plantarum (MCIMB 8826) L. acidophilus (NR RL-B-4495) and L. reuteri (NR RL-B-14171) B. longum

Alginate with a multilayer chitosan coating Alginate or alginate-chitosan

Extrusion

Improved cell viability (8 log CFU/mL) after 4 weeks of storage time Improved cell viability after 39 days (6.5 log CFU/mL for both probiotic bacteria). Control cells are below detection limit after 20 days Improved cell viability (>5.5 log CFU/mL after 6 weeks of storage) Improved cell viability using alginate beads coated with chitosan compared to alginate

Krasaekoopt and Watcharapoka, 2014 Tootoonchi et al., 2015 Bhat et al., 2015

Eleutherine americana extract, oligosaccharides extract, and commercial fructo-oligosaccharides mixture Alginate

Extrusion

Improved cell viability, enhanced sensory characteristics

Phoem et al., 2015

Extrusion

Improved cell viability

Gaanappriya et al., 2013

Orange juice Peach juice

L. paracasei L26

Orange juice

L. rhamnosus GG and L. acidophilus NCFM L. acidophilus 5 and L. casei 01

Orange juice

Orange juice Orange juice Pomegranate juice Pomegranate juice Peach nectar

Pineapple juice

Sapodilla juice Watermelon juice

L. acidophilus

Extrusion

Nualkaekul et al., 2012 García-Ceja et al., 2015

220  Chapter 6  Role of Encapsulation in Functional Beverages

have added beads to a pasteurized longan juice and measured the effect of the combination juice/probiotic on the gut microbiota, using the simulator of the human intestinal microbial ecosystem (SHIME). They found that fatty acid concentration (acetate, propionate, and butyrate) was increased. Moreover, Gaudreau et  al. (2016) co-encapsulated Lactobacillus helveticus and green tea extract in calcium pectinate microparticles. They have observed that co-encapsulation of bacteria with green tea extract provided an additional protection to the cells in gastric conditions. They have suggested that whey protein-coated pectinate microparticles could be a new carrier for the combined delivery of viable probiotic cells and green tea extract to the lower part of the GIT. Prasanna and Charalampopoulos (2018) examined to microencapsulate probiotic bacteria Bifidobacterium longum subsp. infantis CCUG 52486 using the extrusion method in different matrices namely sodium alginate-cow milk (SACM), sodium alginate-casein hydrolysate (SACH), SA, and sodium alginate-goat milk (SAGM) to check the survival of free and encapsulated bacterial cells under various conditions. The encapsulation surface, size, and yield morphology of the microcapsules were analyzed. The survival of microencapsulated bacterial cells and free bacterial cells was evaluated under simulated gastrointestinal conditions as well as in refrigeration, goat milk and cow milk during storage at 4°C for 28 days. Cow milk and goat milk-based matrices resulted in thick microcapsules, which let to improved performances in simulated gastrointestinal conditions than SACH and SA microcapsules. The bacterial cells encapsulated in SAGM demonstrated the highest survival rate in cow milk (7.61 log cfu/g) and goat milk (8.10 log cfu/g) after the storage of 28 days. The cells encapsulated in SACH and SA and the free cells performed poorly under simulated gastrointestinal circumstances and in all various storage conditions. This study showed that SAGM and SACM are appropriate to encapsulate Bifidobacterium longum subsp. infantis CCUG 52486 using the extrusion technique and more precisely, SAGM has a potential to be used as a new encapsulation material for encapsulating probiotic bacteria, resulting in milk and goat milk-based products with higher probiotic cell concentrations during refrigerated storage (Prasanna and Charalampopoulos, 2018). Furthermore, Lactobacillus rhamnosus GG was spray dried in matrices containing whey proteins and various starch ratios. These formulations were placed into apple juice and stored (at 4°C and 25°C) for 5 weeks. It was observed that all formulations containing whey proteins alone or in combination with starch provided better protection to Lactobacillus rhamnosus GG in apple juice or citrate buffer compared to the formulation containing starch alone. The ability of whey proteins to create a buffered environment within the particle

Chapter 6  Role of Encapsulation in Functional Beverages   221

may explain this result. In these conditions, the bacteria are segregated from the stresses of the low pH of apple juice (Ying et al., 2013). Lactobacillus rhamnosus GG was also protected by droplet extrusion in alginate and stored in two fruit juices (pH 2.4 over 28 days). After storage, free cells showed complete probiotic mortality, while beads enhanced probiotic viability after juice storage (Doherty et al., 2012). Besides, Gandomi et  al. (2016) microencapsulated Lactobacillus rhamnosus GG and found 4.5 times higher than control bacteria count after 90 days in apple juice. Antunes et  al. (2013) encapsulated Bifidobacterium animalis subsp. lactis BB-12 using spray drying methods. The spray-dried bacteria count was found to be 8 log CFU/mL after 30 days in acerola nectar, whereas the control bacteria count was found to be 5.9 log CFU/ mL. Petreska-Ivanovska et al. (2014) microencapsulated Lactobacillus casei 01 using spray drying method through polymer complexation and cross-linking with calcium. As wall material, they have used ­chitosan-Ca-alginate enriched with fructooligosaccharide. They have found out that microencapsulated Lactobacillus casei 01 bacteria has an enhanced viability (8.50 log CFU/mL) compared to control (5.70 log CFU/mL) after 6 weeks of storage. Fratianni et al. (2014) encapsulated Saccharomyces cerevisiae var. boulardii using extrusion method with alginate-inulin xanthan gum in cowberry juice and they have observed enhanced viability of S. cerevi­ siae var. boulardii after fermentation and storage. Moreover, Nualkaekul et  al. (2013) encapsulated Lactobacillus plantarium (NCIMB 8826) and Bifidobacterium longum (NCIMB 8809) using the extrusion method with alginate or peçtin with a gelatin, chitosan, or glucomannan coating. They have added into cranberry juice and observed that the double gelatin-coated beads have the highest viability after 6 weeks of storage. Besides, Krasaekoopt and Kitsawad (2010) and Wang et  al. (2015) have studied the encapsulated probiotic microorganisms in grape juice. Krasaekoopt and Kitsawad (2010) have used extrusion method to encapsulate L. casei 01 with alginate-chitosan coating. They have observed high viability of probiotic bacteria. Besides, Wang et  al. (2015) encapsulated Bifidobacterium adolescentis using water-in-emulsion in pea protein-alginate mixture and have observed that at Bifidobacterium adolescentis have higher survival rate at 22°C during storage. Furthermore, there are many recent studies on encapsulation of probiotic bacteria in orange juice (Tootoonchi et al., 2015; Rodrigues et al., 2012; Krasaekoopt and Watcharapoka, 2014; Sohail et al., 2012; Bhat et al., 2015; Wang et al., 2015). All these studies resulted in the increased viability of tested probiotic bacteria. Other studies performed in pomegranate juice (Nualkaekul et al., 2012; Doherty et al.,

222  Chapter 6  Role of Encapsulation in Functional Beverages

2012; Bhat et  al., 2015) also confirm that encapsulation of probiotic bacteria increases their survival rate. In addition, Phoem et al. (2015) studied in pineapple juice and observed improved cell viability of Bifidobacterium longum encapsulated using extrusion method with various wall matrices including oligosaccharides extract, Eleutherine americana extract, and commercial fructooligosaccharides mixture. Gaanappriya et  al. (2013) also encapsulated Lactobacillus acidophi­ lus using exrusion method with alginate wall material and observed improved survival of Lactobacillus acidophilus in sapodilla juice and watermelon juice.

6.4.6 Increasing Diversity of Beverages 6.4.6.1  Color and Flavor (Long-Lasting/Slow-Releasing and Masking Off Flavors) The release of aroma compounds should be controlled according to aroma perception of functional beverages (Ramaekers et al., 2014). The slow release of aroma compounds results in a weak rapid aroma perception, whereas a quick perception leads to a brief burnt flavor resulting in unbalanced flavor profile of functional beverages (Charles et  al., 2015; Guichard et  al., 2013). The role of encapsulation is to provide a controlled release of an encapsulated aroma at acceptable rate during processing, storage, and consumption. Cyclodextrins are used in this purpose for ensuring controlled flavor release. Encapsulation of aroma compound can also decrease the intensity of off-flavors through contact between aroma and oxygen ions and also avoiding direct exposure to light (Kfoury et al., 2016). Tamamoto et  al. (2010) observed that the addition of cyclodextrin decreased the bitterness intensity of ginseng about 50% in a model energy drink. Encapsulation also increases the stability of a wide range of color pigments in functional beverages. Kim et  al. (2014) studied lycopene nanoemulsions for beverage applications and observed that degradation of lycopene nanoemulsions was stable at both 4°C and 20°C. Mainly, phenolic compounds can restrain the off-flavor development in ultrahigh temperature (UHT) treated milk, but little has been investigated for lipophilic phenolic mixtures that are to be encapsulated for even distribution in milk. Guan and Zhong (2017) investigated the physicochemical properties of ferulic acid ethyl ester (FAEE) encapsulated in sodium caseinate and the inhibition of volatile formation after UHT processing. It was observed that encapsulated FAEE was stable after heating at 138°C for 16 min and UV radiation at 365 nm for 32 h. The encapsulated FAEE at a level of 0.18–1.42 mg/mL

Chapter 6  Role of Encapsulation in Functional Beverages   223

suppressed the formation of 2-acetyl-2-thiazoline in model UHT milk by 32.8%–63.2% after 30-day storage at 30°C. Therefore, they have ­suggested using FAEE encapsulated in caseinate to enhance the quality of UHT milk (Guan and Zhong, 2017). Moreover, encapsulation can be used to suppress undesirable flavor and aroma resulted from probiotic bacteria. Sohail et  al. (2012) encapsulated Lactobacillus rhamnosus GG and Lactobacillus acidoph­ ilus NCFM using impinging method and alginate as a coating material in orange juice. They have observed that encapsulation of probiotic bacteria reduced the acidification and negative sensory effects of probiotic bacteria in orange juice (Sohail et  al., 2012). Comparable results were presented by Gandomi et  al. (2016) in apple juice, and by Krasaekoopt and Kitsawad (2010) in orange and grape juices and Phoem et al. (2015) in pineapple juice. Nevertheless, precipitated cells in peach nectar gave an unpleasant display and flavor (fermented) (García-Ceja et al., 2015).

6.4.6.2 Foam-Producing Gas-infusing or turbulence-inducing microparticles have been produced to provide a foamy texture to beverages like instant cappuccino and other coffee mixes, instant refreshing beverage mixes, instant milkshake mixes (Perez and Gaonkar, 2014).

6.4.6.3 Clarification The general purpose of the juice clarification is to reduce the amount of phenolic compounds and decrease the astringency of the product (Alper and Acar, 2004). Ultrafiltration is a method applied to reduce the phenolic compounds, however, it has some drawbacks such as increasing haze in the juice according to the reactions of phenolic compound that cannot be retained by this method. Therefore, an enzymatic pretreatment technique, hyperoxidation of raw juice with laccase (Maier et al., 1994) and pectinase (Lieu Abdullah et al., 2007) before ultrafiltration has been introduced as an alternative method. Gassara-Chatti et al. (2013) studied to produce thermal stable hydrogel formulations of ligninolytic enzymes from Phanerochaete chrysosporium that are more effective on juice clarification. They have observed that the polyphenolic decrease and clarity improvement in mixed juice of berry and pomegranate was important (P > .05) using encapsulated enzymes treatment than free enzymes (Gassara-Chatti et al., 2013). The breakdown of polysaccharides including cellulose and peçtin is an important procedure in fruit juice processing for clarification. Irshad et al. (2017) immobilized pectin lyase (PL), polygalacturonase (PG), and pectin methylesterase (PME) using several concentrations

224  Chapter 6  Role of Encapsulation in Functional Beverages

of chitosan and dextran polyaldehyde as a macromolecular cross-­ linking agent. They have observed significant improvement in the thermal profiles and pH after immobilization of these enzymes. Improved clarification effect was performed in mango, peach, apple, and apricot juice clarification when they were immobilized using chitosan (Irshad et al., 2017). Recently, Shahrestani et  al. (2016) synthesized 1,3,5-triazine-­ functionalized silica encapsulated magnetic nanoparticles (MNPs) and observed its effects on fruit juice clarification. They have used various analytical tools such as transmission electron microscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, and X-ray powder diffraction methods in order to analyze structure, morphology, and properties of functionalized NPs. As a result, Xy-MNPs had important effect on juice clarification and performed significant stability, even after 10 reaction cycles in the enrichment of the juices clarification as it could still maintain about 55% of the initial activity. The Xy-MNPs performed similar clarity enrichment for three types of fruit juices, after 5 h incubation at 50°C. They have also investigated the effect of metal ions and organic solvents on the activity of immobilized xylanase. The results showed an increase in the catalytic activity of Xy-MNPs in the presence of some metal ions while they inhibit the activity of free xylanase. Moreover, in the presence 50% (v/v) organic solvents, both free and immobilized xylanase were inhibited, whereas the Xy-MNPs activity was significantly increased in the presence of 10% (v/v) organic solvents (Shahrestani et al., 2016).

6.4.6.4  Controlled Release Encapsulation methods can be used favorably in controlled release of some compounds in functional beverage products. Zhang and Zhong (2018) studied solid/oil/water (S/O/W) emulsions as delivery systems with retained lactase in milk and controlled release during in  vitro digestion for lactose-intolerant people. Spray-dried lactase powder was suspended in anhydrous milk fat/Span 80 emulsified by sodium caseinate and lecithin (5:1). The S/O/W emulsion had an encapsulation efficiency of 75%, a hydrodynamic diameter of 292 nm, and a zeta potential of −17.37 mV. Cross-linking the dialyzed emulsion with transglutaminase eliminated the detection of free lactase after freeze drying emulsions and the addition of sodium caseinate further preserved lactase activity. The hydrolysis of lactose in full-fat or skim milk after 3-week storage reduced from >75% for free lactase to <15% for encapsulated lactase. The encapsulated lactase was released gradually during the simulated digestions to hydrolyze lactose in milk more efficiently than free lactase. They have suggested S/O/W

Chapter 6  Role of Encapsulation in Functional Beverages   225

emulsions are potential delivery systems to incorporate lactase in milk products (Zhang and Zhong, 2018).

6.5 Conclusions There is a growing interest of consumers in beverages with health-promoting and value-added encapsulation systems that are able to deliver functional bioactive compounds. Increased bioavailability and targeted delivery of these bioactive compounds are of great interest. There is current development in nutrigenomics and metabolomics in designing micro- and nanotechnologies that able to deliver bioactive compounds through foods, tailored to individual genetic prototypes, according to specific metabolic requirements. However, there is still important to understand the complex physicochemical and physiological mechanism involved when added compounds and juices pass through the gastrointestinal system. Besides, consumers are also interested in natural and healthy foods and beverages that have functional properties. Natural beverages will greatly benefit from microencapsulation innovations that able to protect natural and less processed ingredients through specialized physical encapsulation techniques. In addition to traditional microencapsulation technologies such as spray drying, spray chilling, hot melting, extrusion, and coacervation, other newer technologies such as PEGylation, nanoemulsions, and electrospinning for the generation of nanofibers and nanotubes are gaining interest, which might play an important role in future innovations of commercial relevance in the functional beverage industry. Encapsulation provides great potential to protect valuable compounds such as unsaturated lipids, phenolic compounds, vitamins, and probiotics from undesirable effects of environmental conditions during their storage. They also enhance their bioavailability and bioactivity. Encapsulation technology is important for creativity in the functional beverage market according to their functions in color and flavor of beverages. They can be used in the controlled release of aroma, clarification of fruit juices, foaming, enhancing aroma and flavor, and improving the color of beverages. In the near future, functional beverage producers will increasingly face the challenge of having to produce novel products to address consumer demands and the ever more aggressive role of supermarkets producing their own brands. Encapsulation technologies will continue in manufacturing creative, diverse, and value-added products and will have an important part in areas such as food safety, nutrigenomics, and metabolomics.

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Yoon, Y., Choe, E., 2009. Lipid oxidation and stability of tocopherols and phospholipids in soy-added fried products during storage in the dark. Food Sci. Biotechnol. 18, 356–361. Zhang, Y., Zhong, Q., 2018. Freeze-dried capsules prepared from emulsions with encapsulated lactase as a potential delivery system to control lactose hydrolysis in milk. Food Chem. 241, 397–402. Zokti, J., Sham Baharin, B., Mohammed, A., Abas, F., 2016. Green tea leaves extract: ­microencapsulation, physicochemical and storage stability study. Molecules 21, 940.

Further Reading Doherty, S.B., Gee, V.L., Ross, R.P., Stanton, C., Fitzgerald, G.F., Brodkorb, A., 2011. Development and characterisation of whey protein micro-beads as potential matrices for probiotic protection. Food Hydrocoll. 25, 1604–1617. Rocha, M.A.M., Coimbra, M.A., Nunes, C., 2017. Applications of chitosan and their derivatives in beverages: a critical review. Curr. Opin. Food Sci. 15, 61–69.