Fortification in Beverages

FORTIFICATION IN BEVERAGES

3

Asif Ahmad⁎, Zaheer Ahmed† ⁎

Department of Food Technology, PMAS-Arid Agriculture University Rawalpindi, Rawalpindi, Pakistan †Department of Home & Health Sciences, Allama Iqbal Open University Islamabad, Islamabad, Pakistan

3.1  Introducing Food Fortification Concept Food fortification is a common practice to improve the nutrients status in a food product that helps people to combat deficiency symptoms for various ailments. A similar term that often used interchangeably with fortification is enrichment that refers to the addition of those nutrients in the food that lost during the processing operation. Both processes use macronutrients or micronutrients in the food to enhance the nutritional status. Basic objective for both applications is to reduce the population sufferings about nutrient deficiencies. To add a specific nutrient in the food, often staple food in a specific region is selected that may include, cereals, condiments, salts, milk, flour, and some other commonly used food items and the product is developed by the inclusion of micronutrient or macronutrient in that food. At global level, World Health Organization (WHO) also defined the fortification in a similar way as "the practice of deliberately increasing the content of an essential micronutrient, that is, vitamins and minerals (including trace elements) in a food irrespective of whether the nutrients were present originally in the food before processing or not, to improve the nutritional quality of the food supply and to provide a public health benefit with minimal risk to health." The codex definition for food fortification also focuses on these concepts “the addition of one or more essential nutrients to a food whether or not it is normally contained in the food for the purpose of preventing or correcting a demonstrated deficiency of one or more nutrients.” Another well-recognized definition for fortification explains this process as the addition of nutrients or nonnutrient bioactive components to food products to cure or prevent prevalent nutrient insufficiencies by balancing the total nutrient value of diets, to bring back nutrients lost during processing of foods, or to make foodstuffs more attractive to customers. Certain nutrients are added back to foods which were lost earlier during processing through enrichment process, for example, refined grains with Production and Management of Beverages. https://doi.org/10.1016/B978-0-12-815260-7.00003-1 © 2019 Elsevier Inc. All rights reserved.

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iron, thiamine, riboflavin, and niacin, in which nutrients lost during processing are added back. In other cases, certain nutrients are added in foods which are not usually present, for instance, the addition of calcium to orange juice. Although some foods are fortified or enriched according to Food and Drug Administration (FDA) regulations and requirements, certain foods are fortified with nutrients as per producer’s preference. Food and Agriculture Organization (FAO) along with WHO is striving hard to lessen the nutrient deficiencies in masses from all over the world. They have introduced many fortification programs in various countries with the approval of their respective governments. Their activities on food fortification and food enrichment saving millions of lives across the globe each year (FAO, 2014; Dwyer et al., 2015; Edmondson, 2016; Mahan and Raymond, 2016). A wide range of products can be developed using diversified fortification substances and fortification techniques. Among beverages, several classes of fortified products are already available in the market that is expected to grow in future. This may include fortified fruit-based drinks and juices, vegetable-based fortified beverages, dairy-based fortified beverages, citrus fortified beverages, chocolate-based fortified beverages, powdered fortified products, malt-based fortified beverage products, beans-based fortified beverages, nectars, instant flavored fortified beverages, fortified sports drinks, special fortified beverages for pregnant women. These products are available in all parts of the world and their consumption is increasing over the period of time. These products are either fortified with single nutrient or technology of multiple nutrient fortifications may be adopted (Arif et al., 2012; Ahmed et al., 2008; Barclay, 1998). Most of the diet-related nutrients deficiencies prevail in low­income countries characterized by high population. These countries have larger population suffers from iron deficiency anemia, goiter, osteoporosis, stunted growth, and some other physiological problems. It is estimated that more than 50% of pregnant women in these regions suffers from anemia. For low age children, the prevalence of anemia is about 40%, which shows a low iron intake in these two groups. Vitamin D is one of the major deficient vitamins in children and women, its deficiency can be marked with low levels of 25 (OH)D and causes problems related to bones and teeth (Palacios and Gonzalez, 2014). These organs are also affected by insufficient intake of calcium, it not only causes the fragility of bones in adults but its deficiency is also associated with colon cancer and hypertension. The leading causes of osteoporosis and osteomalacia also relate to acute calcium deficiencies in the body (Nemati et al., 2016; Roberts and Stein, 2017). Underdeveloped countries also have a great deficiency of protein intake in their daily life. The less intake of protein also associated with deficiencies of some other micronutrients. These micronutrients are:

Chapter 3  Fortification in Beverages   87

vitamin A, zinc, iron, and folates. It is estimated that one billion people on this globe are deficient in protein intakes. This means most of these people may have deficiency symptoms for zinc, iron, folate, and vitamin A (Wu et al., 2014; Byrd-Bredbenner et al., 2014). Some countries have a mandatory requirement for fortification; United States and Canada have documented rules and regulations for fortifications of many micro and macronutrients in the form of vitamins and minerals. Both countries have a great history of fortification of common salt with iodine to address the problem of goiter. In South East Asia, Pakistan has a well-regularized program for iodine fortification of salt. Most of the countries on this globe targeted rickets problem using vitamin D fortification. The examples of Canada are in citations for rickets and deaths due to vitamin D deficiencies but now their government has addressed these problems through vitamin D fortification programs (Rosenberg et al., 2004).

3.2  Diet-Related Deficiencies Problems Micronutrients are minerals and vitamins that consumed in very small amount but are essential for biochemical processes in the body. Micronutrients deficiencies for vitamin A, iodine, and iron are of greatest concern worldwide for WHO and other international agencies (WHO, 2014). Thirty countries reported that their population was affected by iodine deficiency in the year 2012 (Zimmermann and Andersson, 2012). Iodine deficiency associated with goiter, thyroid function abnormalities, and brain damage. Vitamin A also an essential vitamin for body functioning, its deficiency cause night blindness and xerophthalmia in children (Zayed et al., 2015). Prevalence of iron deficiency for a longer time causing anemia (Alaofè et al., 2017), the deficiency also lead to impaired mental health, impaired physical development in children and premature death (Wong et  al., 2014). Thiamine deficiency is responsible for beriberi. Low-level vitamin D linked to impaired endothelial function, higher coronary artery calcium scores, increased vascular stiffness and inflammation (Kunadian et al., 2014). Vitamin D also plays a vital role in calcium uptake for osteoporosis prevention, rickets prevention, and bones health (Cosman et al., 2014). The diets deficient in B-group vitamins have various problems as vitamin B1 deficiency causes beriberi (Barennes et al., 2015), niacin deficiency cause pellagra (Shi et al., 2017), folate-deficient diet in pregnant women leads to a birth defect in infants (Harika et  al., 2017). Vitamin K and zinc responsible for protein synthesis, however, the deficiency of zinc cause growth retardation, cognitive impairment, and cell-mediated immune dysfunction (Prasad, 2013). Vitamin K is a clotting factor that helps blood clotting when cause injuries but the

88  Chapter 3  Fortification in Beverages

deficiency of vitamin K cause more bleeding in young infants (Schulte et al., 2014). Selenium is the only mineral that acts as an antioxidant help to protect from free radicals. Keshan, a disease is associated with selenium deficiency (Loscalzo, 2014). Low calcium intakes found in many developing countries that may lead to calcium deficiency in the human population. The osteomalacia, rickets, and bone diseases recognized to be associated with calcium deficiency (Shin et al., 2015). Some of the diseases or conditions in the human body can be reversed using macro and micronutrients through fortified food and beverage products. These diseases/conditions appear due to insufficient intake of certain nutrients, Table 3.1 summarized various disease caused by insufficient intake of nutrients.

Table 3.1  Deficiency Symptoms or Disease in Relation to Nutrients Nutrients

Deficiency Symptoms/Disease

References

Protein

Low energy level, sluggish, weak muscle mass, marasmus, Kwashiorkor, weak immune system Night blindness, xerophthalmia, bitot spot, keratomalacia Fractures, colorectal disease, cognitive decline, depressed mood, autoimmune thyroid disease Cystic fibrosis, truncal and limb ataxia, ptosis, spinocerebellar ataxia, ophthalmoplegia, muscle weakness, and dysarthria Deficiency is very uncommon but symptoms include, difficulty in clotting, bleeding, new-borns may have vitamin K deficiency, warfarin therapy, adjunct treatment for osteoporosis Wernicke encephalopathy, korsakoff syndrome, poor memory, muscle cramps Fatigue, anaemia, nerve damage, mouth and lip sores, sluggishness, sore throat Pellagra, inflammation, skin problems

Geissler and Powers (2017) and Hujoel and Lingström (2017) LeBlanc et al. (2015) and Johnson (2014)

Vitamin A Vitamin D Vitamin E

Vitamin K

Vitamin B1 (Thiamine) Vitamin B2 Vitamin B3 Vitamin B6 Vitamin B12

Mouth and eye problems, hair loss, inflammation, dermatitis Periodontitis, weakness, tiredness, pale skin, intestinal problem, nerve problems

LeBlanc et al. (2015), Theodoratou et al. (2014), and Cummings et al. (2016) Niki and Traber (2012), Johnson (2014), and Hall (2015) Schwalfenberg (2017) and Machlin (1991)

Abdou and Hazell (2015) and Hall (2015) Byrd-Bredbenner et al. (2014), Machlin (1991), and Csapó et al. (2017) Hall (2015), Machlin (1991), and Csapó et al. (2017) Byrd-Bredbenner et al. (2014) and Csapó et al. (2017) Zong et al. (2016), Brito et al. (2016), Stabler (2013), and Moll and Davis (2017)

Chapter 3  Fortification in Beverages   89

Table 3.1  Deficiency Symptoms or Disease in Relation to Nutrients —cont’d Nutrients

Deficiency Symptoms/Disease

References

Vitamin C

Scurvy, bruising, swollen gums, delayed would healing, rough skin, scale formation, gingivitis, weak immune system Lethargy, fatigue, breath shortness, irritability, pale skin Anaemia, fatigue, glossitis, oesophageal webs, dysphagia, restless legs, cognitive impairment Weak and brittle bones, weak nails, depression, numbness, muscle spasms, hallucinations, memory loss, muscle cramps, osteoporosis Muscle weakness, stupor, coma, weak bones, irritability, numbness, low appetite, impaired tooth development Feel of cold, lethargies, slow mental response, unusual weight gain, goiter, poor memory, difficulty in concentration, muscular fatigue Deficiency is un common. Weakness, nausea, confusion Tingling, nausea, vomiting, palpitation, constipation, weakness, abdominal cramps, psychosis depression Keshan disease, selenoproteins

Hall (2015), Machlin (1991), and Buckley et al. (2014)

Folates Iron Calcium

Phosphorus

Iodine

Sodium Potassium

Selenium

Byrd-Bredbenner et al. (2014), Reynolds (2014), and Moll and Davis (2017) Hall (2015) and Moll and Davis (2017) Byrd-Bredbenner et al. (2014), Prasad (2013), and Aslam and Varani (2016) Hall (2015), Machlin (1991), and Neufeld et al. (2017) Byrd-Bredbenner et al. (2014) and Zimmermann and Boelaert (2015) Prasad (2013) and Pohl et al. (2013) Machlin (1991), Byrd-Bredbenner et al. (2014), and Pohl et al. (2013) Prasad (2013) and Loscalzo (2014)

3.3  Need for Food Fortification As per above discussions on diet-related nutrient deficiencies and focus of WHO, FAO, and Codex for fortification programs there is a great need for food fortification to save millions of people all over the world. There are several areas that require special attention to food fortification programs. Some of the agricultural and processing practices cause some losses of nutrients that are often referred to as good manufacturing practices, good agricultural practices, and good handling practices and good storage practices at field and industry level. The addition of nutrients at these stages is essentially required to restore the original nutritional status of the food product that is

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­ resent in edible parts of that food. This requires the knowledge about p the original nutrients available within the food items and every effort must have focused to restore those nutrients through fortification program that are lost during storage, handling, and processing (Stahl, 2014). A good example that is practiced in several parts of the world is adding vitamin B1 (Thiamine) in cereal flour in the amounts that were lost during handling and processing to restore actual amounts as present in the raw material. These fortification techniques are also applicable in beverages that may have natural minerals in it or can be fortified with essential minerals provided that water of higher purity is used (Mehmood et al., 2013; Khalid et al., 2013). Another purpose of food fortification is to achieve the equivalence of substituted food. Margarine and butter are almost the same food (substituted food), butter is made from natural animal’s fats while margarine is considered as a good substitute for butter but made from a plant source. Butter naturally contains good amounts of vitamins A, D, and E. Therefore, the substituted product (such as margarine) should be fortified with same levels of vitamin A, D, and E as present in butter. Vitamin D is required in an amount of 10–20 μg/day that seems to be the very low amount. Even though most of the people are not getting the required amount of vitamin D, this condition requires the needs for viable policies for vitamin D fortification at the national level in the countries facing such problems. Protein is one of the abundant nutrients available through plant and animal foods but FAO depicts a very alarming situation for protein intake across the globe. It is estimated that about one billion people are deprived of the recommended intake of protein. The challenge to combat protein deficiency situation will be on the rise with the growth of global population and requires special fortification programs to fight against its deficiency (Cashman and Kiely, 2016; Wu et al., 2014; UN-FAO, 2013). There are numerous health problems associated with insufficient intake of vitamins and minerals including bone health, night blindness, scurvy, and some others. The solution of these common problems lies in efficient food fortification programs. Fortification to be more effective, it is necessary to add diversified minerals and vitamins in food and beverage in appropriate amounts that match recommended daily allowances. Selection of appropriate raw material is also important for fortification into a specific product, for instance, there are more than 10 calcium salts that can be added to milk-based beverages. To combat micronutrient and macronutrient deficiencies in children, WHO have its recommendations for nonbreast fed as well as breastfed children of age 6–23 months. These recommendations suggest the use of fortification materials with higher bioavailability (Gibson et al., 2015). Diet-related deficiency symptoms never appear individually, in most of the cases these appeared in combination. For

Chapter 3  Fortification in Beverages   91

instance, protein deficiencies symptoms appear along with zinc, vitamin, and some other nutrients deficiency symptoms. Similarly, bone weakness symptoms are not only due to deficiency of calcium but it also shows the deficiencies of phosphorus and vitamin D. Therefore, to handle these problems there is a need to develop a strategy of multiple nutrients fortification in a single product and beverages that may provide a viable solution to the problem. The knowledge about characteristics of each nutrient added into a beverage is key to success in multiple nutrient fortification programs. For instance, if a processor intends the iron fortification in a beverage product, the sole use of iron salt for fortification will have limited bioavailability. Its bioavailability can be improved through the addition of ascorbic acid or some other organic acid that will enhance its absorption by making suitable chelates in the body (Barclay, 1998). Keeping in view the large segment of the people, in various parts of the world, is suffering from macro and micronutrient deficiencies. Therefore, every country in the world should make national level policies in conjunction with global policies for food fortification. These policies at country level should always be supported by some renowned global organizations such as WHO, FAO, UNICEF, the US Centres for Disease Control and Prevention (CDC), the Global Alliance for Improved Nutrition (GAIN), and Nutrition International (Wu et al., 2014; UN-FAO, 2013; Jefferds et al., 2013). There is a range of motives and benefits that provide motivation for beverage fortification. Consumer demand is one of the most important reasons behind this action. Consumers are very conscious about the nutrients that they are taken through the beverage products and these beverage products are consumed by every segment of the population. Based on this idea processors are encouraged to develop fortified products according to needs of the consumer. As the needs and demands of the consumer are highly variable, therefore, industrialists must produce a diversified form of fortified beverages to satisfy every segment of the population. Sometimes, a diet fortified beverage suffices the need of the consumer but some time high energy drink with macro or micronutrients addition is indisputably required for health-conscious consumer, another consumer may require these essential nutrients in the simple treated bottled water.

3.4  Fortification in Food and Beverage Products Beverage consumption is a part of every society and culture having an image of healthy foods and drinks. Consumers always want healthy dairy-based beverages, soymilk, mineral water with added

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nutrients, yogurt-based beverages, and much more similar kinds of the products. With developing beverage industry, processors bring new nutritional fortified products to capture the market as per the demand of the consumer. Consumer is not only conscious about flavors but also mindful of health-imparting nutrients. Knowledge of fortification strategies enables industrialists to meet the consumer demands for healthy alternatives. Surveys in the United States and other advanced countries indicated a high demand from the consumer for micronutrients fortified beverages. This showed a great prospect for developing novel fortified beverage products on part of industrialists (Hexagon_Nutrition, 2015). Fortified nutrients in these beverages may be classified as micronutrients and macronutrients and may include several forms of vitamins, minerals, and certain bioactive substances. A brief representation of these nutrients that can be fortified in beverage products is shown in Fig. 3.1. The stability of these fortification substances may vary and depend on several factors. Among these, the vitamins stability influenced by moisture, oxidizing agents, temperature, antioxidants, reducing agents, oxygen, pH, light, and minerals. Processing conditions and storage of beverage products may also affect their presence. Among water-soluble vitamins, B1, B12, and ascorbic acid are more susceptible to losses but among fat-soluble vitamins the most susceptible vitamin is D. Losses in these vitamins may accelerate due to poor storage and packaging conditions, oxidative changes

Iron

Folic acid (vitamin B9)

Niacin

Calcium

Selenium

Iodine

Zinc

Vitamin D

Probiotic

Fortification in beverages

Vitamin B6

Dietary fiber

Plant sterol

Vitamin B12 Bioactive peptides

Riboflavin (vitamin B2) Vitamin A

Bioactive peptides

Omega-3 fatty acid

Fig. 3.1  Summary of different fortified substances in beverages.

Chapter 3  Fortification in Beverages   93

during storage also reduce their activities. These losses demand a viable fortification mechanism that may ensure high levels of these nutrients in the beverage product for a longer period of time (Barclay and Haschke, 2015). Beverages are good vehicles for the addition of nutrients so that it can efficiently transport nutrients to various parts of the body through its consumption along with meal (Pszczola, 1998). England has a great history to launch a beverage in 1980s with the name of Aqua Libra. According to the label, the ingredients include the juice of fruits like grapes, apple, carbonated spring water, grapes, flavors, and extract of sunflower seeds, sesame seed, and tarragon. Many such other products are also available in markets over the year with addressing health problems. Japanese pharmaceutical company developed a soft drink containing dietary fibers, vitamins, and minerals which marked as a bestselling drink in Japan. Coca-Cola Company also developed their fiber-rich beverage named as Fibi. A drink named as Zija fortified with the leaves of Moringa oleifera which is a great source of antioxidants, omega oil, minerals, and vital proteins (Ansari and Kumar, 2012). Here are details of some beverage products fortified with micro and macronutrients:

3.4.1 Calcium Fortification in Beverage Calcium (Ca) have a multidimensional role in the body, the major role is to provide strength and structural edifice to the bones and teeth and used in their developmental phases. Apart from bones and teeth, calcium has numerous physiological roles in the body. Its presence is associated with coagulation of blood during bleeding; it has a physiological role in muscle contraction, cell division during mitosis and facilitates the nerve transmission process (Nemati et  al., 2016). Comparing other nutrients, the body needs for calcium is slightly higher. More awareness about bone health in relation to calcium for prevention of osteoporosis is the deal of interest today. Several bioavailable forms of calcium exist in nature and food processors make use of these forms in various food products for food fortification. Most used forms are calcium carbonate, calcium citrate, calcium malate, calcium lactate, and calcium glycerol phosphate. Among these and several others, those forms are preferred for fortification having greater bioavailability. For fortified beverages, choice of calcium salt is driven by its organoleptic properties. The color and flavor in a beverage are most important sensory parameters. Therefore, added salt of calcium should have bland taste, white or colorless coloring properties and it should not impart off flavor to the beverage. The calcium content in available salts is also highly variable and range from 9% to 71% as in gluconate and oxide form, respectively. A brief description of calcium salts with their characteristics is presented in Table 3.2.

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Table 3.2  Selective Calcium Salts Used for Fortification Calcium Salt

Chemical Formula

Calcium Content (%)

Calcium carbonate

CaCO3

40

Calcium sulfate Calcium chloride

CaSO4

29

CaCl2

36

Ca-citrate

Ca3(C6H5O7)2

24

Tricalcium citrate

(C6H5O7)2Ca3 4H2O

21

Calcium lactate

C6H10CaO6

13

Calcium lactate gluconate

C9H16CaO10

11–12

Tricalcium phosphate

Ca3(PO4)2

38

Characteristics

References

Odorless salt; have water solubility of 0.013 g/L; have good stabilizing properties in beverages Odorless; have good water solubility of 0.21 g/100 mL Colorless crystalline solid; have solubility in CH3COOH, alcohols and water; good water solubility of 74.5 g/100 mL (20°C) Water solubility 0.95 g/L (25°C); impart buffering action in beverage, acid regulator White, odorless, crystalline powder or fine powder; have fair solubility in water and acidic media; have better solubility at lower temperatures Solubility 6.7–9.74 g/dL; solubility increase as gluconate ions increase, white or off white powder Used in effervescent mixes and beverages; neutral taste; solubility in water 22 g/100 mL; exists as mixture of calcium lactate and calcium gluconate It is a white solid of low solubilitysolubility in water 0.002 g/100 g

Lehmann and Joseph (2015) Allen et al. (2006) Gimeno et al. (1998)

Sakhaee and Pak (2013) Sakhaee and Pak (2013)

Nemati et al. (2016)

Trailokya et al. (2017)

Trailokya et al. (2017) and Nascimento de Paula et al. (2014)

Naturally, dairy-based products including fresh milk are a good source of calcium. As concerned fortification, calcium salt addition in dairy-based beverages is considered as a better choice. Beverage products require more solubility of calcium salt fortification material, this lead to the inclusion of calcium gluconate, calcium citrate, and calcium malate in fortified beverages. However, calcium carbonate, calcium sulfate, calcium chloride, calcium acetate is preferred for baked items fortification. In milk-based liquid beverages fortification, tribasic calcium phosphate is one of the salts of choice. The

Chapter 3  Fortification in Beverages   95

c­ omposition of milk some time causes sedimentation of this fortified calcium salt. This problem can be tackled by use of appropriate gum or hydrocolloids. These fortification materials can also be used to fortify yogurt drinks with the addition of suitable hydrocolloids (Deeth and Lewis, 2015). In milk, the presence of lactose causes indigestion problems in some individuals. To replace natural animal milk, processors introduced plant-based soymilk. This milk product is now available in industrialized countries and high-income countries and can be consumed in many ways as a beverage. New experimentations on soymilk beverage are under exploration. This also includes fortifying soy beverage with appropriate calcium salts. Calcium gluconate is considered one of the appropriate calcium salts that can be used in soy beverage for fortification purposes. Another calcium salt is potassium lacto gluconate, but there are some limitations with these salts most important is their low level of calcium contents, others are undesirable changes in color and texture as soy protein cross-linked with proteins (Allen et al., 2009). Calcium lactate gluconate is highly recommended calcium salts due to its most soluble nature, additionally, least affects the other physical properties of beverages (Mohammadi et al., 2016). This calcium salts nowadays substance of choice for fortifying drinking water providing a satisfactory bioavailability of calcium in aqueous drinking systems in a manner similar to that in dairy and other pharmaceutical supplements (Cotruvo, 2006). This calcium fortified drinking water imparts a beneficial impact on human health by preventing bones disorders. Research is also in progress to explore utilization of calcium salts in fruit and vegetable drinks. Consumption of excess calcium through beverages was not reported as a health issue. However, the addition of calcium in the clear beverages may cause undesirable flavor and destabilization of proteins (Singh et al., 2015).

3.4.2 Iron Fortification in Beverage   There are numerous dimensions of iron in the body to regulate metabolic processes. Iron in many forms plays a vital role as components of certain metabolic enzymes that is, cytochrome system that involved in oxidative metabolism. In the human body, iron act as oxygen carrier from lungs to tissues usually found as part of hemoglobin. The deficiency of iron is common nutritional disorder problem throughout the world. Iron imbalance for the long term may cause anemia that defined as low blood hemoglobin concentration. Other major links are also associated with iron deficiencies including impaired cognitive behavior, effects on productivity and physical performance during pregnancy that is leading cause of anemia. The major population is affected by iron deficiency including pregnant women, children, and infants. About 20% of maternal motility and low birth weight account

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due to iron deficiency however, in case of acute anemic conditions, the chances for child and mother mortality may increase (Brabin et al., 2001a,b; Kozuki et al., 2012). Briefly, selective signs and symptoms for iron deficiency are presented in Fig.  3.2. There are numerous documented risk factors associated with iron deficiency; these risk factors serve as guidelines for nutritionists to determine the extent of iron deficiency. Iron-fortified foods and beverages can reverse the adverse effects of iron deficiency. Consumption of iron-fortified beverages during childbearing days have a positive impact to lessen mortality chances in mother and infants, similar results can be obtained by using iron supplements, fortified baked items and other iron-fortified products (Cogswell et al., 2003; Ahmed et al., 2014). As, iron supplementation in form of syrup and tablets failed due to side effects of high concentration of iron found in syrup and tablets. Therefore, fortification is most effective to reduce iron deficiency prevalence (Gabriel et al., 2005). Various iron salts having low price and high bioavailability are used; among these salt FeSO4 is more beneficial for fortification. For water-based beverages ferrous gluconate used due to its good solubility. Other useful salts that may be used for fortification purposes are: ferrous fumarate, ferrous tartrate, ferrous succinate, and ferrous citrate. In addition to these salts, ferrous amino acid chelates along with previously mentioned salts can be used as a mixture to attain higher bioavailability during fortification process (Mohammadi et  al., 2016). Selection of suitable salt based on its solubility can be fortified in water and sports drinks. Iron considered to more challenging micronutrients to add to food and beverages because it has the best bioavailability that interacts with food constituents to produce undesirable changes. If the iron-fortified food with an elevated level of the unsaturated fatty acid system is stored for prolong time, it can cause rancidity and subsequently produce off flavor. This happened due to prooxidant properties of iron that facilitate oxidation process in unsaturated fat containing liquid beverages. Similarly, calcium addition in iron-fortified food and beverages may influence its bioavailability negatively. To overcome these problems, either level of iron salt is increased or some organic acid such as ascorbic acid is added along with iron salt in the beverage product (Walczyk et al., 2014). Another strategy to improve iron status in human is to improve hemoglobin concentration; the technology of multimicronutrient fortification in beverage product has a great application. Multimicronutrient fortified juices when consumed by school going children in high doses, moderate doses, or low doses resulted in increased hemoglobin levels to a significant value. This resulted in significant decrease of anemia in tested subjects (n = 246). It is suggested that school feeding programs may effectively use multimicronutrient fortified beverages to overcome the problem of anemia and certain other nutrient deficiency problems (Angeles-Agdeppa et al., 2017).

Chapter 3  Fortification in Beverages   97

Signs and symptoms of

iron deficiency Fatigue and tiredness

Restless leg syndrome

Increased sensitivity to cold

Shortness breath

Depression Frequent headache

Hairloss

Brittle nails

Fig. 3.2  Pictorial representation of signs and symptoms of iron deficiency.

Iron fortification is perhaps the most difficult task in beverage products. A good knowledge of iron salts for their characteristics is required to choose appropriate material for fortification. Most of the iron salts, although have good water solubility yet addition into aqueous system, leads to unacceptable flavor and color. High level of technological knowledge and skills are required to overcome these problems.

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The selection of suitable iron salt based on solubility and organoleptic properties is challenging for beverage processor. From a nutrition point of view, the bioavailability of fortification salt about inhibitors is also among the major issues for fortification purposes. The presence of some natural inhibitors in fruit and vegetable may reduce absorption of iron salt in the human body, that is, very challenging for researchers working in the field of food science and technology. On basis of bioavailability, ferrous sulfate is preferred among various iron salts used in fortification process, but it has limited utility in milk-based beverages as it can interact with unsaturated fats of milk causing oxidation reactions to provoke rancidity and off flavors in the beverage products (Hurrell, 2002). For milk-based beverages or infant formulas, ferric pyrophosphate as a fortification material has greater application. However, it has lesser bioavailability as compared to ferrous sulfate. One of the advantages of using this salt is its complimentary effects in chocolate-based milk drinks to develop iron fortified chocolate beverages, this drink resulted in a moderate level of bio-absorption of iron into the human body, this absorption has a time dependency and takes slightly higher time to absorb. To enhance further the absorption process, slightly higher amounts are recommended for the fortification of ferric pyrophosphate. Recently, a new ferrous containing salt namely ferrous ammonium phosphate was tested in the dairy beverage product. This salt was compared with ferrous sulfate and ferric pyrophosphate. Ferrous sulfate showed highest iron absorption followed by ferrous ammonium phosphate, as the ferrous sulfate have some stability issues in full cream milk thus ferrous ammonium sulfate has a greater potential for fortification in milk-based beverages to achieve higher bioavailability targets (Hurrell et al., 1991; Walczyk et al., 2013). In a research trial, some infants with a low socioeconomic group from Pakistani population were fed wheat-milk beverage samples containing ferric pyrophosphate or ferrous fumarate with extra amounts of 3–5 mg/day. This fortification strategy resulted in significant increase in hemoglobin and serum ferritin in these children (Glinz et al., 2017; Hurrell, 2002).

3.4.3  Zinc Fortification Zinc plays a vital role in cellular growth, cell division, and protein synthesis; it is also an essential component of a large number of enzymes. All the population may have a risk of zinc deficiency but ­children and infants are more susceptible. The pregnant women are also vulnerable to zinc deficiency (Sian et  al., 2002; Osendarp et  al., 2003). Worldwide the deficiency in zinc is not well documented. However, FAO estimated that 1/5th population on this globe is at risk for its deficiency. Identification of zinc deficiency is hard to detect in moderate and mild deficiency. However, severe deficiency shows the

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symptoms of dermatitis, diarrhea, growth retardation, and mental disturbance. Some of the signs and symptoms of zinc deficiency are presented in Fig. 3.3. The major risk factors for deficiency of zinc include a diet low in zinc, impaired utilization of zinc, malabsorption disorders, and other genetic diseases. Zinc supplementation in children improved growth rates and reduced infectious diseases, diarrhea, and pneumonia. Severe deficiency of zinc in pregnant women shows poor maternal pregnancy outcomes and low birth weight of infants (Mohammadi et al., 2016; Khalid et al., 2014; FAO, 2014). The zinc deficiency can be prevented by increasing the intake of zinc fortified beverages. Zinc fortification is not very easy. Zinc compounds that are added to beverages are gluconate, picolinate, chloride, acetate, citrate, ascorbate, zinc oxide, sulfate, and zinc amino acid chelate (Mohammadi et al., 2016). Zinc gluconate and zinc amino acid chelate are mostly preferred over other. To reduce the iron deficiency formulation of fruit flavor dry beverages of zinc and iron supplement with or without vitamins being focused. However, the ­addition of ­water-soluble iron compounds with zinc salts may impart flavor changes and produced a metallic taste. But water fortifies with at least 5 ppm zinc and 1 ppm iron in form or amino acid chelates shows no flavor and metallic taste (Mehansho et al., 2002).

3.4.4 Selenium Fortification in Beverage Selenium is vital mineral required for healthy functioning of the body, its antioxidant activity is very much like the vitamin E activity and has a sparing effect of vitamin E. Usually, it acts as an antioxidant

Signs and symptoms of

zinc deficiency Zinc is a important trace mineral which is useful for the body in many ways. It is essential for cell division & aids normal growth and development during pregnancy, childhood and adolescence. Co

mp im rom mu ise nit d y

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Fig. 3.3  Pictorial representation of signs and symptoms of zinc deficiency.

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to protect against free radical and have health implications in heart disease, arthritis, and cancer. For beverage fortification, this mineral is added in sports drinks and mineral water along with several other micronutrients in a way to follow the concept of multiple micronutrient fortifications (Mehansho et al., 2002). Apart from sports drinks and bottled mineral water, selenium has a great potential to be added in dairy-based beverage product to develop functional beverages. The French market has now introduced dairy beverage products with added selenium content that have a great synergy with magnesium as fortifying material. Some research from New Zealand focused on improving selenium content in cow and sheep milk (Özer and Kirmaci, 2010). Some researchers introduced the idea of vital minerals and selenium in tea that have an influence on physiochemical as well sensory properties of the tea beverages. Tea samples were fortified at the level of 2.2 mg/kg, in addition to these samples tea also contain citric acid and powdered sugar (Adnan et al., 2013; Zhang et al., 2014).

3.4.5 Iodine Fortification in Beverage Iodine deficiency in the past was perhaps the most prevailing nutrient deficiency in most parts of the world. Its deficiency was estimated to a value of 29% in world population and associated with goiter and lower mental capabilities. The problem is more aggravated in school going children resulted in reduced ability to learn during schooling period (Black et al., 2013). Its deficiency can be controlled using sole fortification of iodine source such as in table salt or it can be fortified as multiple micronutrient fortifications of beverages. Although multiple micronutrient fortifications for iodine are very limited, they can serve as a useful technique to cover the gap for its deficiency. The advantages of using beverages as a fortification vector are due to higher consumer acceptance and flexible delivery as ready to consume beverage products (Aaron et al., 2015). A study was carried out to evaluate the effects of multiple micronutrient fortified beverages on various parameters in school children. The fortified and nonfortified beverage was administrated to school going children for 16 weeks. The multiple nutrient fortified beverages resulted in the betterment of anemic conditions. It was observed that in groups of children that were administrated with iodine fortified beverages caused an increase in urinary iodine excretion. Iodine-deficient subjects when consumed the iodine fortified beverage showed significant signs of improvement in the fitness test and cognitive performance. Clinical trials also confirmed the efficiency of multiple micronutrients fortified beverages to combat against micronutrient deficiencies (Solon et al., 2003).

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3.4.6  Folic Acid (Vitamin B9) Fortification in Beverage Folic acid also is known as vitamin B9 or folate plays an important role in the formation of nucleic acid that intervenes in tissue growth and cell multiplication. According to British Dietetic Association (BDA) folic acid helpful for brain health in infants, hearing losses, and creating more blood cells. Folic acid is one of the most common micronutrients that deficient in pregnant women, its deficiency in women causes neural tube and childbirth defects along with brain and spine damage to newborn baby (Harika et al., 2017). The pregnant women should, therefore, consume enough folic acid that helps prevent the fetus from developing deformities of brain and spine. Pregnant women should take either supplement or consumed folic acid rich diets. For pregnant women, the experts recommend 400 μg of folic acid per day from the beginning of pregnancy. Green leafy vegetables, fruits, and yeasts are the main sources of folate. Taking these foods and cereal grains in low amounts than recommended may increase the risk factors for folic acid deficiency. Evidence found that iron with folic acid is better in preventing anemia than iron alone (Allen and Casterline-Sabel, 2001). Effective programs for folic acid fortification lower plasma homocysteine in population and prevention of neural tube defects even having a low prevalence of folic acid deficiency. Researchers estimated that folic acid fortification prevented spina bifida by 25% globally in 2012 (Youngblood et al., 2013). Folic acid is a water-soluble vitamin, having good stability in aqueous systems thus can be effectively used in beverages for fortification. Not only the beverages but a variety of other food products can be fortified using a folic acid such as energy bars, drinks, and ready to eat cereals (Yang et al., 2010). However, Food and Agricultural Organization and WHO support fortification of folic acid into cereal flours (Allen et al., 2006). The exceed level of folic acid may mask vitamin B12 deficiency. In recent years, a fermented milk beverage was developed along with cereals and selective strains having the potential to produce folate. Despite adding folate in the beverage, this research used microorganisms to produce folate as a strategy to add folate to barley fermented milk, corn fermented milk, and wheat fermented milk. Encapsulated strains of Lactobacillus plantarum and Streptococcus thermophilus were used for this purpose during the fermentation process. From a total of six tested beverage sample, corn fermented milk using L. plantarum and wheat fermented milk using S. thermophilus depicted a great folic acid production potential in the beverage product. That ranged between 0.295 and 0.349 mg/L (Sharaf et al., 2015). In a similar attempt, Deep et al. (2017) used the strategy of probiotics fortification in addition to folic acid in the beverage product. This

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strategy ensures a high level of folates with the favorable component matrix in the beverage product. The researchers claimed that tomato juice and orange juice with a high level of folate contents can be produced using S. thermophilus NCIM 2904 strains. This strategy also ensured the stability of folate content in the beverage product. A recent study from Guatemala comprising of 236 women and 156 children were provided beverage products fortified with folates and other nutrients. Consumption of folate-fortified beverage influences positively the body status of folate in these subjects (Oliva et al., 2016).

3.4.7 Thiamine (Vitamin B1) Fortification Thiamine plays a critical role in energy metabolism and also in growth, development, and function of cells (Coates et  al., 2010). Thiamine from foods or supplement is absorbed by the small intestine. The excess thiamine stored in liver but it has very short life. So, continuous consumption of thiamine from the diet is thus necessary. The thiamine deficiency is a public health problem in many parts of the world. Severe deficiency of vitamin B1 leads to beriberi. This disease usually appears due to consumption of high carbohydrate foods (rice) mainly in form of polished rice and intakes of raw fish containing anti thiamine compounds. Wheat germs, yeast extract, and green vegetables are main sources of thiamine. The risk factors for thiamine deficiency is consumption of diets that contain antithiamine compounds for example, raw fish contains thiaminase (Lonsdale, 1990). The severe deficiency of thiamine leads two distinct forms of beriberi known as dry beriberi and wet beriberi. The dry beriberi associated with chronic and neurological while wet beriberi associated with fatal heart failure. Several studies indicate that supplementation can reverse the symptoms of thiamine deficiency. Sulfur dioxide has a detrimental effect on thiamine which results in thiamine degradation. As some fruit juice contain sulfur dioxide so prior to fortification the level of SO2 should be determined. The multimicronutrient strategy can be used as potential fortification mechanism to develop beverages with sufficient amount of thiamine (Yang and Huffman, 2011).

3.4.8 Riboflavin (Vitamin B2) Fortification in Beverage To add riboflavin in beverage product a different approach was used in a recent research attempt. The fermented milk-based beverage was produced by adding cereal flours and selective strains of microorganisms meant for the fermentation process. These microorganisms have a capacity to develop riboflavin in the beverage product. For fermentation, strains of L. plantarum and S. thermophilus were used. Wheat milk, fermented

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by L. plantarum and barley milk fermented by S. thermophilus; the research was carried out in selective village population of Guatemala. The beverage product was prepared using riboflavin and some other nutrients were distributed among the village population. The beverage fermented with S. thermophilus showed a great potential to produce riboflavin in the resultant beverage products to a level 2.12–1.82 mg/L (Sharaf et  al., 2015). In another research, beverage products fortified with riboflavin was made available to selective subjects among village population. The beverage consuming groups comprised of females and school going children. Some subjects were provided riboflavin fortified beverages in 350 mL bottles and other was provided with nonfortified beverages in the same volume bottles. The corresponding sampling from these individuals indicated a great contrast between fortified and nonfortified groups, a variation in an increase of riboflavin was observed in both groups with a value of +16.4 vs +5 (Oliva et al., 2016). A study conducted in Botswana for multinutrient fortified beverages indicated a great potential of these products to eradicate micronutrient deficiencies in developing countries. In this study, 311 school going children were selected and provided 240 mL serving of either experimental fortified beverage containing about 12 micronutrients or placebo drink with the same energy content as the experimental beverage. The study was conducted for 2 months. This study improved the riboflavin status in experimental group and improvement was reported in weight for age and upper arm circumference of the children that were provided experimental multinutrient fortified beverage samples (Abrams et al., 2003). To evaluate the effective fortification and retention of riboflavin in beverage samples over a period of time, its evaluation methods should be well defined. Recently, a new user friendly method for riboflavin determination in alcoholic products was reported. This method ensures accuracy for determination of riboflavin in alcoholic beverage products and can be adopted at the industrial level. The method is based on fluorescence quenching effect using riboflavin binding protein. This method also involves the use of single-diode fluorimeter. This fluorometric method is comparable with traditional HPLC method of riboflavin determination. It is even considered better for performance and much cheaper as compared to traditional HPLC method; these qualities make it more agreeable for the alcoholic beverage industry. A similar situation exists for riboflavin determination in cola beverage that is considered cumbersome at the industrial level. This analysis situation becomes more time consuming and difficult if the cola beverages contain caffeine and caramel along with riboflavin fortification. Therefore, a comparatively rapid method for the industry was developed, which simultaneously determines the caffeine, caramel, and fortified riboflavin in cola beverages. The method is equally good for energy drinks in which riboflavin is added through ­fortification.

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This new method is based on recording synchronous fluorescence spectra at a wavelength ranging from 200 to 500 nm. Detection process involved the use of HPLC system with fluorescence detection. High coefficient of determination as obtained during this method development indicates the effectiveness of this method for simultaneous determination of riboflavin along with other substances. The new model is highly effective and is comparable to traditional used HPLC method. The data were confirmed using 20 cola drinks and 16 sports drink samples of different brands. Research indicated good statistical characteristics based on calibration and prediction data. Overall, this new method is highly valuable in saving resources in terms of cost and time and has a great potential to adopt for relevant beverage industry (Bonamore et al., 2016; Májek et al., 2014).

3.4.9 Niacin (Vitamin B3) Fortified Beverage Niacin is a micronutrient having several regulatory functions, and most important regulatory function shows its role in oxidative and reduction reaction in the metabolism of nutrients. Deficiency of niacin is associated with skin disease commonly known as pellagra. High prevalence of this vitamin exists in pregnant and lactating women in all parts of the world but Mali, Burkina Faso, Mozambique, Philippines, and Bangladesh are the most affected countries and need special fortification strategies to overcome the problem. Bangladesh introduced fortified beverages containing about 5 mg niacin in a single serving, which corresponds to 31% of recommended nutrition intake on daily basis (Hyder et al., 2007; Yang and Huffman, 2011). Adopting multinutrient fortifying strategy, niacin when fortified with other nutrients and administrated to women before gestation, it resulted in higher mean birth weight (Yang and Huffman, 2011). Niacin fortification can be practiced in synergy with zinc, iron, copper, selenium, iodine, and certain w ­ atersoluble vitamins. However, in fortified beverage products nutrients such as phosphorus, calcium, and potassium may influence the organoleptic characteristics in a negative way (Hofmeyr et al., 2010).

3.4.10  Pyridoxine (Vitamin B6) Fortification in Beverage Vitamin B6 is a group of three naturally occurring compounds pyridoxine, pyridoxal, and pyridoxamine. The vitamin B6 acts as a coenzyme of various enzymes that are involved in amino acid metabolism. The body helps to make several neurotransmitters that carry signals from one nerve to another. It maintains the health of red blood cells, immune system, and nervous system and also helps in the breakdown of protein in the body. The deficiency of vitamin B6 seems relatively

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uncommon but mildly deficient shown in few studies. The deficiency symptoms include irritability, anxiety, muscle pain, fatigue. The study shows that deficiency of vitamin B6 found in people having Alzheimer’s disease (Malouf and Grimley Evans, 2003). Although the deficiency of vitamin B6 is not common, however, elderly people in which energy requirement is low and they are feeding on low energy nutrient, they can catch up vitamin B6 deficiency. A Dutch survey of elderly people within the age limit of 65–80 years indicated a vitamin B6 deficiency in 54% subjects. To treat this deficiency, fortification with vitamin B6 is a better and easy strategy. Nutrient dense food is recommended for this purpose that may include some of the beverage products (Berendsen et al., 2016).

3.4.11 Vitamin B12 Fortification in Beverage Vitamin B12 is an important water-soluble vitamin having vital functioning for brain and nervous system. This vitamin is rarely added alone during fortification process. Most often this vitamin is added as multimicronutrient fortification strategy with other micronutrients in the beverage and some other food products. In beverage products, vitamin B12 addition has a good synergy with vitamin D, folates, and riboflavin. When beverage products are developed using these nutrients, it is tested on rural population facing these nutrient deficiencies. In addition to the fortification, micronutrients influence positively various biomarkers associated with these micronutrients including vitamin B12. Some of the signs and symptoms of deficiency diseases in response to less intake of vitamin B12 are shown in Fig. 3.4. Baseline to end line data showed a positive increase of 46.2% vitamin B12 versus a value of 6.2% in fortification vs non fortification groups (Oliva et al., 2016). A different strategy for addition of vitamin B12 is recently reported by some investigators. The researchers coated the ready-toblend fruit and vegetable salad with vitamin B12 and chitosan ate at the rate of 0.25 mg/L and 10 g/L followed by a storage period of 9 days at refrigerated temperature (5°C). This stored material was used for beverage preparation at 0, 4, 7, and 9 days. Coating of chitosan ensures avoidance of undesirable changes due to chemical reaction or microflora. Fortified beverage products have good levels (8.6 μg/kg) of vitamin B12 on processing day and even after 9 days storage. It is recommended that consuming 200 mL of this beverage can provide recommended a daily intake of vitamin B12 through this strategy and can be stored at refrigerated temperature (Artés-Hernández et al., 2017).

3.4.12 Vitamin A Fortification in Beverage Vitamin A is an essential vitamin for normal functioning of the visual system, immune functions, maintenance function of cell, reproduction, and integrity of epithelial cells. However, it required in small

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Vitamin B12 deficiency

Bleeding gums Hypothyroidism

Depression

Low energy

Cognitive decline Numbness

Daughter

Son

Who are they? Fig. 3.4  Pictorial representation of signs and symptoms of vitamin B-12 deficiency.

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amounts. Normally, vitamin A found in wide range of foods usually food of vegetable and fruit origins. Changes in vision and eyes, specifically night blindness and xerophthalmia are traditionally relied on the deficiency of vitamin A (Zayed et al., 2015). About 3 million of preschool children have an ocular sign of vitamin A deficiency approximately half of which died within a year after blindness. The deficiency also associated with measles and diarrhea in children. The common risk factors for vitamin A deficiency are poor nutritional status, consumption of vitamin A-deficient diets. The supplementation or fortification of vitamin A in food and beverages can reduce the adverse effects. Fruits and vegetables are a great source of vitamin A usually contain carotenoids, β-carotene is the most important. Beverage fortified with β-carotene not only serves as a precursor of vitamin A but it also provides color to the beverage. Vitamin A for skim and reduced fat milk required minimum 2000 international unit for fortification (Yeh et al., 2017). Vitamin A is light sensitive and it degrades when exposure to light. The wide acceptance of vitamin D fortification in milk led to the fortification of vitamin A in milk that was initiated in the 1940s. Vitamin A is fat soluble vitamin which found in significant amount in whole milk but fat removal in fat-free milk and low-fat milk reduced vitamin A, therefore, fortification of vitamin A usually done in these kinds of milk. Vitamin A fortified in the form of retinyl palmitate, which is an ester of retinol and palmitic acid in fluid milk. The vitamin A found in milk as in natural form tends to more stable in light than added vitamins. So, the stability of added vitamin A may affect by light, heats, acids, and ultraviolet radiation (Yeh et al., 2017). The addition of vitamins to milk can influence the milk flavor that may due to enzymatic degradation of milk fat and proteins. Immediately after processing milk pasteurization imparts cooked flavor (Boelrijk et al., 2003). Vitamin A fortified usually in skim and low-fat milk imparted detectable off flavor. Sensory experiments thus needed for vitamin A-fortified milk to remove this problem. Fortification of vitamin A in any kind of food and beverages having more moisture level tends to adverse effects on the stability of vitamin A. However, to overcome this problem encapsulated fortification techniques with additional moisture barrier was evaluated. Vitamin A fortification is limited to milk and milk products. Carotenoid can be also used as a source of Vitamin A (Whited et al., 2002).

3.4.13 Vitamin D Fortification in Beverage Vitamin D although required in very low amount but have significance important for many physiological reasons. The average daily intake is 10–20 μg that approximately corresponds to 400–800 IU/ day. The variability in amounts is dependent on physical conditions, growth, age, sex, and exposure to sunlight. National levels survey in some countries reported low levels intake of vitamin D; these ­situations

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need to develop fortification strategies. Vitamin D is soluble in fatbased systems; therefore, fortification of this nutrient requires a media containing fats in it. The deficiencies are usually due to limited exposure to sunlight, low intake of milk, or consuming vitamin D-deficient foods, and decrease in cutaneous synthesis. Consequences of all these may lead to limited bones health-osteomalacia in adults and rickets in children. Some other skeletal effects are also found related to vitamin D. Vitamin D reduces the risk of coronary heart diseases and cancer (Chowdhury et al., 2014). Mostly foods are low in vitamin D so to achieve the recommended intakes and other public health strategies fortification of vitamin D are increased. Usually, milk and milk products are fortified with vitamin D. The functional beverages are fortified by two forms of vitamin D ergocalciferol (D2) and cholecalciferol (D3). When the beverages are fortified with these vitamins, the types of vitamins must list in ingredients statements of products. Vitamin D mostly used for fortification of fluid milk as it is also recommended by dietary guidelines advisory committee (DGAC, 2015). The initial fortification of vitamin D began in the 1930s from United State where the vitamin was fortified in fluid milk to reduce rickets in children. Pasteurized milk required to be fortified vitamin D at a minimum of 400 international units the acceptable level is 400–600 IU per quart of milk (Yeh et al., 2017). The vitamin concentrate is used to fortifying milk added before pasteurization. The acceptable fortification concentrations of vitamin D were 80%–120% that is specified by the FDA in Pasteurized Milk Ordinance (PMO). The over fortification of vitamins A and D leads to intoxication, kidney failure, and soft tissue damages (Jacobus et al., 1992). Vitamin D added in the form of D3 a synthetic vitamin in fluid milk. This synthetic vitamin made from irradiation of animal fats mainly from the waxy secretion from sheep skins that called lanolin (Holick, 2005). For dairy processing systems, there is two forms of vitamin premises; oil based and water based. So the oil-based added to milk after cream separation and water-based premix added before separation. The addition of vitamin in milk usually takes place after separation and before pasteurization, and then homogenization takes place for uniform distribution of vitamin. The vitamins are added to milk by using either metering pump procedure or batch procedure so both procedures required accurate measurement of vitamin concentration (Yeh et al., 2017).

3.5  Fortification in Dairy-Based Beverages Dairy foods including beverages contribute ~52% of the calcium, 52% of the vitamin D, 27% of the vitamin A, 26% of the phosphorus, 25% of the vitamin B12, 26% of saturated fat, 14% of total fat, 10% of

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sodium, and 10% of total calories to the diets of American adult (Cifelli et al., 2017). Apart from the well-established function of improvement of bone health, more recent evidence suggests that an intake of milk and dairy products is associated with lower risk of obesity, cardiovascular disease, and type 2 diabetes (Thorning et al., 2016). Dairy-based beverages can be fortified with antioxidants, ω-3 fatty acids, vegetable proteins due to their higher consumer acceptance and the requirement for simple storage conditions at refrigerated storage (Corbo et al., 2014) and stabilizers play an important role to maintain the desirable rheology of these dairy products (Ali et al., 2000; Tasneem et al., 2014). There is a growing interest of consumers in dairy-based beverages that have been fortified with omega-3 fatty acids. Different variants of dairy beverages with added omega-3 fatty acids have been reported that may include: ultrahigh-temperature milk (UHT), fresh milk, ice cream, yogurt, and milk-based drinks. Nestle world-renowned dairy producer brand started Omega Plus line in South America and for the Far Eastern markets (Panse and Phalke, 2016). Liquid yogurt and yogurt drinks are also included among famous and most consumed dairy beverage all over the world. Owing to growing trend of nutraceuticals market; yogurt has been fortified with the number of fortification materials and bioactive substances. Yoghurt has been fortified with a variety of minerals and fat-soluble vitamins (A, D, E, and K) have been reported to provide strength against various body problems and facilitate various processing operation for improved quality during the manufacturing of the product at the industrial level. On the other side, most of the water-soluble vitamins did not tolerate the high processing temperature of dairy products and get lost from the product (Gahruie et al., 2015). Coculturing of common yogurt strains with L. kefiranofaciens is another way to enhance the nutritional and functional properties of the dairy-based drinks (Ahmed et al., 2013). Deficiency of vitamin D may lead to osteoporosis, rickets especially in childhood, type-I diabetes, and multiple cancers including colon cancer. Owing to this yogurt-based beverage products are good candidates to be fortified with vitamin D; therefore, such beverage products are extensively reported in the scientific literature in reference to fortification (Wagner et al., 2008; Kazmi et al., 2007; Gahruie et al., 2015). To some extent, vitamin A and vitamin C also have a potential in dairy beverages but too little is found in the scientific literature. Vitamin A is added as carotenoids or it may be added as retinol acetate and retinol palmitate. Some selective carotenoids with intact β-ionone ring act as a precursor for vitamin A and also provide color to beverage product. Choice of carotenoids is important, carotenoids with higher biological value and color-imparting properties are selected for that purposes to have an appropriate color shade of the beverage along with the nutritive value (Preedy et al., 2013; Gahruie et al., 2015).

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3.6  Fortification Using Bioactives There is a new trend in beverage manufacturing that involves strategies to add bioactive substances. These bioactive substances provide additional health benefits to fight against certain diseases.

3.6.1  Probiotic Fortification Probiotic dairy products are a significant class of functional as well as fortified foods. Dairy-based beverage products are mainly fermented products account for around 43% of the practical beverage marketplace in five key European markets, United States, Japan, and Australia (Mikkola and Colantuono, 2017). Probiotic fortification is frequent in beverages as they are considered outstanding vehicles for probiotics microorganisms (Gürakan et al., 2009). Presently probiotics are defined as administration of live microorganisms which present a health benefit to the host on administration in adequate amounts (Butel, 2014). The health benefits attributed to probiotics are diversified; for example, improvement in lactose-intolerance symptoms, lowering of hypertension, lessening of symptoms aroused through antibiotic cure of Helicobacter pylori, management of colon cancer symptoms, alleviation of atopic eczema among infants, improvement of symptoms of Crohn’s disease, urogenital health care and enhancement of immune function, and provision of macro and micronutrients (Iqbal et al., 2014). Due to numerous benefits on human health, probiotic bacteria have become the focus of attention of food industries, especially dairy industry (Grover et al., 2012). For that reason, an increasing number of dairy manufacturers are adding Lactobacillus species and Bifidobacterium species to some of their beverage products. The most commonly included probiotic bacteria among lactobacilli include L. acidophilus, L. casei, and L. rhamnosus and Bifidobacterium bifidum among bifidobacteria. A few examples of beverage products incorporated with L. casei are Actimel (Danone, France), Immunitas, and Yakult (Yakult Honsha Co, Japan) while L. johnsonii and L. helveticus in Shirota, and Chamytor (Nestle, France) (Corbo et al., 2014). The success of probiotics in dairy drinks is limited owing to restrained environment of the ingredients apprehension over contamination or low viability of strains during storage. The viability of probiotic strains from Lactobacillus spp. and Bifidobacterium spp. in dairy beverages is affected by low pH, the presence of H2O2 and O2, metabolites produced during metabolisms such as lactic acid and acetic acid (Costa et al., 2013; Fonteles et al., 2012).

3.6.2 Omega-3 Fatty Acid Fortification in Beverage Many bioactive components like omega-3 (ω-3) fatty acids are also being added to commercial dairy beverages along with fortification using bioactive components, such as ALA (α-linoleic acid, C18:3n-3), EPA

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(eicosapentaenoic acid, C20:5n-3), and DHA (docosahexaenoic acid, C22:6n-3) (Özer and Kirmaci, 2010). Currently, available ω-3 fortified dairy beverages are drinkable yogurt, milk drinks, and fresh and UHTtreated milk (Shahidi and Ambigaipalan, 2016). In a number of clinical studies ω-3 fatty acids consumption has been found to be associated with proper fetal development, antiinflammatory properties and improved cardiovascular, retinal and immune function, weight management, and treatment of very mild Alzheimer’s disease (Swanson et al., 2012). According to National health and nutrition examination survey, polyunsaturated fatty acid consumption in the United States is <185 mg/day, whereas International Society for the Study of Fatty Acids and Lipids recommends the daily intake of 500 mg/day. By and large, the intake of ω-3 fatty acids is quite low in inhabitants living away from coastal areas (Weylandt et al., 2015). Deficiency of ω-3 fatty acids has been linked with lethargic memory and mental efficiency, tickly feeling of the nerves, reduced vision, increased predisposition of blood clots formation, compromised immunity, increased levels of triacylglycerides and low-density lipoprotein (LDL), damaged membrane functioning, growth retardation in infants and children, hypertension, hyperheartbeat, learning disabilities, and menopausal issues (Shahidi and Ambigaipalan, 2016). There are some undesirable effects of ω-3 fatty acids are reported in the literature, commonly observed undesirable effects of ω-3 fatty acids supplementation include gastrointestinal symptoms such as nausea, discomfort and “fishy” burp (Schuchardt and Hahn, 2013). However, in a large metaanalysis, the favorable risk-to-benefit ratio was found 1:400 (Mozaffarian and Rimm, 2006). Few examples of ω-3 fatty acids fortified products include Natrel Omega-3 (Natrel, Canada) and Heart Plus (PB Food, Australia) (Corbo et al., 2014). Addition of omega-3 fatty acids in food remained a challenge mainly owing to “fishy” taste and flavor which is a basic characteristic of this fortification. Thanks to microencapsulation techniques, which has provided a viable solution to undesirable flavor and now omega-3 fatty acids can be added in multiple beverages and variety of fruit juices using encapsulation technology (Panse and Phalke, 2016).

3.6.3  Bioactive Peptides Fortification Bioactive peptides can be obtained from proteolysis of various proteins by enzymes and also produced during intestinal digestion of nutrients, work as a prospective physiological regulator of metabolism. Recently, great attention has been paid to study the bioactive roles of bioactive peptides as antioxidant, antihypertensive, hypocholesterolemic, antiinflammatory, and immunomodulatory effects (Umayaparvathi et al., 2014; Hernández-Ledesma et al., 2014). Certain food proteins, chiefly milk caseins, may act as a precursor of bioactive

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peptides with varying physiological benefits (Özer and Kirmaci, 2010). An example of commercially available beverage with added bioactive peptides is Evolus from Valio Ltd. (Finland). Biologically active peptides synthesized by L. helveticus from milk casein have the major beneficial effect in it and can be added in beverage products (Prado et al., 2008).

3.6.4  Plant Sterol Fortification Plant sterols (PS) including phytosterols and phytostanols have shown protective effect against cancers of the stomach, lung, ovaries, and breasts (Grattan, 2013) and well-established LDL cholesterol lowering role (Ras et al., 2014). Similar effects can be achieved by flavonoids and phenolic substances, if fortified in food products (Ahmad et  al., 2015). Foods fortified with phytosterols (commonly with the sterol esters) are simply integrated into the fatty part of the product (Duong et al., 2016). The main source of PS for current functional foods and dietary supplements is tall oil, a by-product of the wood pulp industry, and vegetable oil deodorizer distillate (Alemany-Costa et al., 2012). The enrichment of dairy beverages with PS is a favorable way to obtain the daily recommended amount of PS (2 g/day recommended by FDA) to lower cholesterol and in turn reducing cardiovascular disease risk (Nagarajappa and Battula, 2017). Different researchers have reported the potential usage of plant extracts, powder, and fruit juice itself. Anthocyanin and phenolic acids present in plant have established anticancer effect. Different beverages even dairy products especially the yogurt has been fortified with ethanol extract from different grape varieties (Karaaslan et al., 2011; Gahruie et al., 2015).

3.7  Dietary Fiber Fortification in Beverages Rising understanding among the food patrons all over the world regarding the health attributes imparted by fiber-rich diet; attracted the attention of many food scientists. Although dietary fiber did not contribute toward calories, it has numerous physiological impacts on the health of human body. This lead to a great attraction of fiber-based low caloric food products; and such foodstuffs have become an essential component of consumer’s daily diet (Veena et al., 2016). Dietary fiber includes high mass oligosaccharides, polysaccharides, inulin, and prebiotics (Gupta and Sharma, 2016). Dietary fiber addition in beverages has multiple usages both from technological and nutraceuticals point of view. Dietary fiber addition in beverage can act as thickening, gelling, and dispersing and carrying agent. It can also improve the glucose intolerance, lower the blood cholesterol, and decreases the risk of colon and another type of cancers (Hassanpour, 2016; Gupta and

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Sharma, 2016; Maphosa and Jideani, 2016). In recent research, buttermilk which is locally called as lassie has been incorporated into the dietary fiber to increase it nutraceuticals impact. Fiber enrichment in buttermilk noticeably abridged the phase severance in buttermilk (Mudgil and Barak, 2016). Addition of fiber in probiotics-based food enhanced the viability of such strain in beverages containing such hydrocolloids (Gupta and Sharma, 2016). Fiber-based hydrocolloids are typically used at a level of 0.05% and 0.03% in a 250 mL drink; this counts about 0.125 and 0.75 g in the beverage. In the United States to permit a claim, at least 2.5 g per serving is required; whereas in the European Union, its level is 3 g/100 g (Viscione, 2013). Acacia gum is suitable to be used in nondairy beverages since it has the capability to tolerate at low pH. Moreover, it does not lead to enhanced viscosity even at high concentrations; but it imparts mouthfeel impact without masking the flavor release of drinks (Viscione, 2013; Li, 2009; Cherbut et al., 2003). Dietary fiber like polydextrose enhanced the viability of L. acidophilus LA-5 and B. lactis Bb-12 in yogurt-like drink and strains remained stable at storage temperature for 30 days (6 log CFU/mL) (Shori, 2016). In another study, it was depicted that viability of probiotic strain, that is, L. acidophilus LA5 in nonfermented milk carrot; increased to 98%; whereas B. lactis Bb-12, L. plantarum and L. rhamnosus GG depicted viability ranging between 88% and 92% during 20 days of storage at 4°C (Daneshi et al., 2013). Yet in another research, it was depicted that L. casei NRRL B442 in fermented pineapple juice at 31°C 6.03 Log CFU/mL after 42 days of storage at 4°C, that is, refrigerated storage (Costa et al., 2013).

3.8  Consumer Acceptance for Fortified Foods The use of fortified beverages is very common in developed countries. Most of the consumers for these products are children, women, elder chronic patients especially the educated people having good knowledge about fortification. Some groups of the consumer may not necessarily found any nutritional deficiency and considered as a healthy and active people. The publication of US National Health and Nutrition Examination Survey (NHANES) reported that the children 1–13 years and the most senior member of the population were frequent consumers of fortified products (Bailey et al., 2010; Yang et al., 2010). Fortification may influence the consumers purchasing decision. International Food Information Council’s Food and Health Survey’s finding shows that four out of five purchased fortified food and beverages because of its beneficial impacts. One-third believe that fortified products have great influence on overall health (Dwyer et  al., 2014).

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Consumers interest in understanding fortification beverages showing positive response in recent years. Some researchers found that consumer preferred the vitamin D fortified products in Germany. Some other reports depicted the acceptance of biofortified food and beverage products in the large segment of the population. But still, need some awareness about the nutritional and health benefits of fortified foods. This can be done by providing information to the consumers through different sources and by involving media. Beyond, its health benefits fortification is a simple process which is extremely good with its benefit of far outweighing the cost. So, due to its lower costs, the consumer also prefer fortified foods over traditional foods (Birol et al., 2015).

3.9 Conclusion Through fortification process, we can add several nutrients and nonnutrient bioactive components in beverage products. This strategy may be used to overcome the widespread shortfall of nutrients among the large segment of the affected population all over the world. Over the past few decades, fortification appeared as a highly effective technique to reduce the risk of nutrient deficiency diseases such as goiter, beriberi, scurvy, pellagra, and osteoporosis in different parts of the world. But still, along-term strategy is required to combat some other nutrients deficiency diseases across the globe. International agencies working on food, nutrition, and health should combine their efforts with government level agencies in each country to develop fortification program at the regional level. There is a growing trend of consuming fortified food products among the consumers due to growing awareness about its health implications. More exploration is required to characterize the fortification material that may be used for the development of beverage products. We need research-based interventions and diagnosable techniques to understand the effective relationship between fortification and control of nutrient deficiency disease. Combined efforts of various stakeholders are required to make viable policies for the use of the fortified product for a specified market and geographical area. The latest research in the production of fortified beverages, quality control, nutrient-based disease control, and marketing of fortified product is required and the result should be publicly disseminated in all parts of the world.

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