Manufacturing of Funtional Whey Beverage

1. INTRODUCTION

INTRODUCTION

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage

Whey is the yellow, watery liquid that separates from the curd during the cheese making process and contains nearly half of all solids found in whole milk. It is estimated that during the production of one pound of cheese, approximately nine pounds of whey are produced. At one time, this whey was viewed as nothing more than a waste product. Cheese processors disposed of whey down drains until tightened environmental regulations made the dumping process illegal and expensive. Other disposal mechanisms included the discharge of whey into local waterways, the ocean, or as a component in animal feed. Additionally, some whey has also been used as nutrient-laden soil enrichment in a process called land spreading. As land spreading restrictions and water treatment facility regulations continue to tighten over the next few years, cheese manufacturers will be forced to find alternative methods for disposing of or utilizing whey. Drying technologies are available for processing liquid whey into whey protein isolates and concentrates for an abundance of applications, but the energy needs alone can overwhelm small cheese producers. Equipment costs can also be prohibitive. An alternative solution for liquid whey disposal is needed. (Smithers, 2008)

In order to provide assistance to Alma Creamery, the Kansas Department of Commerce, a funding source for this research, has requested the development of a value-added beverage that utilizes unprocessed cheese whey as an alternative to disposing of the whey into the environment. As an extension of previous graduate research by Raymond Kassatly (Kansas State University, Manhattan, KS), the proposed beverage would continue to be formulated with a liquid whey base, but with the added value of carbonation.

1.1. Importance of whey Beverage

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Beverages based on whey continue to receive a considerable amount of attention reflecting a growing awareness of potential of these products in market place. The beverage has high nutritional quality and increased energy value. These could be particularly useful in places where there is lack of food and improper nutrition leading to deficiencies of certain nutrients.

Out of 85 million tones of global production 40% is still disposed as raw whey in to sewage which leads to serious environmental pollution, because of its high BOD of 3, 00,000-5, 00,000ppm. Therefore conversion of whey in fermented and non-fermented beverages is one of the most attractive avenues for utilization of whey for human consumption. (Horton, B.S.1995)

The present investigation has been proposed to develop whey based functional whey beverage by mixing appropriate fruit juice and processed whey with selection of suitable stabilizer ,sugar, colour and flavour to increase the consumer appeal. The fruit like apple, guava, litchi, mango, and pineapple are used for preparation of beverages. (Dr.J.N.de. Wit, 2009)

1.2. Whey Proteins

Whey protein is a protein fraction obtained from cow’s milk. Milk contains two major protein fractions, including casein, which provides about 80% by weight of the total protein, and whey protein, which provides about 20% by weight of the total protein. While its concentration in milk is not significant, whey protein contains all of the essential amino acids, and therefore, is a high quality, “complete source” of protein. More specifically, whey proteins are a rich source of branched chain amino acids (BCAAs), containing the highest known levels of any natural food source. BCAAs are important for athletes, since, unlike the other essential amino acids, they are metabolized directly into muscle tissue and are the first amino acids used during periods of exercise and resistance training. While these nutritional characteristics would benefit athletes, whey protein has the potential to extend its advantages to an average consumer. In a clinical trial presented in 2006 by the United States Department of Agriculture (USDA) researchers found that those “consuming supplemental whey protein weighed less and put on less body fat compared to individuals who consumed a calorie-equivalent carbohydrate supplement” .

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From a functional perspective, dried whey protein is appropriate for beverage formulation in that it has a fresh, neutral taste and, therefore, can be included in other foods without adversely affecting the taste. Any flavor that is imparted from the whey protein lends itself well to citrus and fruit-flavored drinks (Dr.J.N.de Wit, 2009). It is, however, important to consider that unprocessed liquid cheese whey is regarded as nearly unpalatable in its original, unprocessed form. In addition to flavor attributes, whey proteins function in an array of beverages due to their solubility over a wide pH range. The ideal pH of a proposed whey protein beverage would either be far above or below the isoelectric point of whey proteins, which are 4.6. If the beverage is formulated at or near this point, the whey proteins will precipitate and beverage quality and acceptability will suffer. ( Sharma & vahati K.L.,1999)

1.3. Commercial Whey Protein Products

Given the advantages of whey protein, it has become a popular source of nutrition in a variety of forms: whey protein supplement bars, whey protein concentrates, whey protein isolates, and whey protein beverages. According to Mintel International’s Global New Products Database (GNPD), 1,763 products in the United States and 6,435 worldwide were introduced with whey ingredients in 2005. These products require that the whey be processed from its original form through drying technologies, ultrafiltration, and/or hydrolysis treatments. The intent of this research was to utilize the unprocessed cheese whey as the liquid base for a beverage. A broad and informal market evaluation of the current whey protein beverage sector shows no evidence of such a product.

A functional beverage can be defined as a drink product that is non-alcoholic, ready to drink and includes in its formulation non-traditional ingredients. This includes herbs, vitamins, minerals, amino acids or additional raw fruit or vegetable ingredients, so as to provide specific health benefits that go beyond general nutrition. Sports and performance drinks, energy drinks, ready to drink (RTD) teas, enhanced fruit drinks, soy beverages and enhanced water, among others, are some of the product segments rolled out as functional beverages in the market space.

Functional beverages have become popular due to its appeal to consumers who are seeking specific health benefits in their foods and beverages with their

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'healthiness-on-the-go' idea. Both convenience and health have been identified as important factors when consumers make decisions about purchasing foods and beverages. Functional drinks are promoted with benefits such as heart health, improved immunity and digestion, joint health, satiety, and energy-boosting. (Sharma & vahati K.L., 1999)

1.4. Health and wellness

Functional beverage companies are more aware of the ‘health conscious’ individuals and have introduced functional beverages with less sugar and therefore less calories. For example, Vitamin water 10 contains only 10 calories per serving (25 calories for a 351mL bottle, 7.5 grams of sugar and 250% of daily allowance of Vitamin C).On the other hand; it has the same 25% of the daily allowance of Vitamins B3, B5, B6 and B12 as the original. Vitamin water 10 has an all natural sweetener extracted from the stevia plant, which is a benefit in lowering calorie content (although taste is another matter) as well as fitting the product in the "natural" category.

1.5. Weight management

With increased worries about obesity and its implications on health, combined with demand for convenience goods, consumers are naturally looking towards easy weight loss methods that they can easily integrate into their lifestyles. As such, functional beverages are striving to achieve that through addition of ingredients that promote weight loss.

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http://en.wikipedia.org/wiki/Vitamin_C

http://en.wikipedia.org/wiki/Vitaminwater


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Aims and objective


2. Aims and objectives:

The objective of this dissertation was to develop a value-added functional whey beverage that utilizes unprocessed liquid whey as a byproduct of cheese manufacture. Liquid whey accounts for up to ninety percent of the yield during cheese making and has historically been considered a waste product. As legal and environmentally friendly whey disposal options dwindle, alternative utilization strategies for the liquid whey must be developed. Further processing of whey, such as drying, creates a potential financial burden on small cheese producers. Considering the proteins found in whey are associated with known health benefits, the nutrient laden, unprocessed liquid whey is an ideal base for a wholesome beverage. With the advantage of added whey protein and the appeal of carbonation, the resulting carbonated protein beverage would make a unique and successful addition to the growing health beverage sector.

Whey constituted about 80-92% of the volume of milk used for conversion into channa, paneer, cheese and casein. It retained about 45-55% of milk nutrients comprising serum protein, lactose, mineral and vitamins. On an estimate more than 3 million tones of whey is produced in countries while more than 2 lakh tones of it is containing valuable nutrients are dumped into the sewage.

A functional liquid whey based beverage system was developed, keeping in mind the technological and financial capabilities of a small producer. The product was evaluated based on physical, microbiological, and sensory characteristics. With minor formulation changes, this beverage could realistically and competitively exist in today’s marketplace. The following objectives were planned.

1. To develop the functional whey beverages based on whey which is nutritious by product of cheese, channa and paneer industries.

2. To develop a suitable procedure to flavourize the functional whey beverages with the tropical fruits.

3. To identify pro biotic bacteria that could be used as an adjunct culture to improve the therapeutical quality of this dairy beverage for needy people.

4. To study proximate principle of raw materials in whey, fruit juices etc.5. To study the nutritional composition of whey beverages.

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Because of it high organic matter content, milk whey constitutes a serious environmental problem with lactose being mainly responsible for its high BOD and COD values. Acid dairy drinks are worldwide products existing in many variations Ex. Buttermilk, whey drink, kefir etc. This beverage can be described as an acidified protein liquid system with stability and viscosity similar to natural milk. The health benefits provided by probiotic bacteria have led to their increasing use in the fermented dairy products. The lactic acid bacteria occupy a central role in this process and have long and safe history of occupation and consumption in the production of fermented food and beverages. They cause rapid acidification of the raw material through the production of organic acid mainly lactic acid. In this way they enhance shelf life and microbial safety, improve texture and contribute to the pleasant sensory profile to the end product.

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Literature

Survey


3. LITERATURE SURVEY

Growing concern over pollution and environment control has renewed the pressure on the cheese manufacture to stop dumping whey into streams and municipal sewage system. Consequently, the search has begun again for new methods to use whey. In light of growing global food shortages, the most logical use would be to return whey to the human food chain in palatable form. Several authors have suggested that whey could be used in the formulation of nutritious soft drinks or high protein beverages and also might be used as an additive in soaps and fruit juices (Anon 1968, A. G.). Using cheese whey as a beverage in human nutrition, especially for therapeutic purposes, can be traced back to the ancient Greeks; I-Iippoerates, in 460 B.C., prescribed whey for an assortment of human ailment. In the middle ages, whey was recommended by many doctors for various diseases, by the middle 19th century, whey cures reached a high point with the establishment of over 400 whey houses in Western Europe. As late as 1940’s in the spas in Central Europe, dyspepsia, uremia, arthritis, gout, liver diseases, anemia and even tuberculosis were treated with the ingestion up to 1500 g 0f whey per day.

Whey is the by-product of cheese and casein manufacture, being what remains of the milk once the cheese or casein is removed. Generally 100 L of milk produces about 12 kg of cheese or about 3 kg of casein. In either case, about 87 L of whey is made as a by-product.

Whey comprises 80–90% of the total volume of milk entering the process and contains about 50% of the nutrients in the original milk: soluble protein, lactose, vitamins and minerals. Whey as a by-product from the manufacture of hard, semi-hard or soft cheese and rennet casein is known as sweet whey and has a pH of 5.9 – 6.6. Manufacture of mineral-acid precipitated casein yields acid whey with a pH of 4.3 – 4.6. Table 4 below shows approximate composition figures for whey from cheese and casein manufacture.

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Table: 3.1. Approximate composition of Whey

Constituent Cheese Whey % Casein Whey %Total Solid 6.4 6.5Water 93.6 93.5Fat 0.05 0.04True Protein 0.55 0.55NPN(non-protein nitrogen 0.18 0.18Lactose 4.8 4.9Ash (minerals) 0.5 0.8Calcium 0.043 0.12Phosphorus 0.040 0.065Sodium 0.050 0.050Potassium 0.16 0.16Chloride 0.11 0.11Lactic acid 0.05 0.4

Ref: Handbook on whey and whey product by Dr.J.N.de Wit, 2009

Available literature indicates that whey beverage have been studied extensively in Brazil, Australia, Europe, Ger-many, India etc. of published revives of the comprehensive.

We have tried to review all aspects of whey-beverage manufacture which have been developed.

Research at University of Novi Sad, Faculty of Technology, studied that development of whey beverage. In that study considers beverages consisting of whey, fruit components (orange, pear, peach, and apple), citric acid and sucrose. It also offers their optimal composition. The dry matter of the fruit component, the pH of the beverage and its sucrose content were independent variables. Blends were prepared in accordance with the factorial design. After pasteurization, they were exposed to a sensor analysis so that the following characteristics were estimated: flavour, odour, colour, sediment, appearance and total quality (sum of the previous factors). By applying the regression analysis method, a mathematical model of each characteristic was derived. None of the characteristic functions had an extreme, so the maxima lay at the boundaries of the independent variables (6% of dry matter and 4% or 2% of sucrose). Only the pH changed within a narrow range. The statistical method has shown that the quality of blends with orange and pear mostly depends on the sucrose content, while the quality of blends with peach

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and apple depends on the dry matter of the fruit. Interaction of dry matter and sucrose is most significant for the blend with pear, while the balance between sucrose and pH strongly depends on the quality of all the other products. The peach–whey beverage containing 6% of dry matter and 2% of sucrose as well as having pH 3.6 proved to be the best.

Research (Mljekarstvo, 2008) investigates the Whey based beverages-new generation of dairy products. Liquid whey consists of approximately 93% water and contains almost 50% of total solids present in the milk of which lactose is main constituent. Lactose is the main constituent of whey while proteins represent less than 1% of total solids. Minerals and vitamins are present in fewer amounts also. Production of whey based beverages started in 1970's and until today a wide range of different whey based beverages has been developed. They can be produced from native sweet or acid whey, from de proteinised whey, from native whey which was diluted with water, from whey powder or by whey fermentation. Non alcoholic whey beverages include wide range of products obtained by mixing native sweet, diluted or acid whey with different additives like tropical fruits (but also other fruits like apples, pears, strawberries or cranberries), crops and their products (mainly bran), isolates of vegetable proteins, CO2, chocolate, cocoa, vanilla extracts and other aromatizing agents. Special attention is being paid to production of fermented whey beverages with probiotic bacteria where the most important step is the choice of suitable culture of bacteria in order to produce functional beverage with high nutritional value and acceptable sensory characteristics. Non alcoholic whey beverages also include dietetic beverages, drinks with hydrolyzed lactose, milk like drinks and powder drinks. Whey is a very good raw material for production of alcoholic beverages due to the fact that the main constituent of the solid content is lactose (about 70%). Alcoholic whey beverages include drinks with small amount of alcohol (up to 1.5%), whey beer and whey wine. Whey beverages are suitable for wide range of consumers – from children to the elderly ones. They have very high nutritional value and good therapeutic characteristics.

Another research had done in University of Novi Sad, Faculty of Technology, and Navi Sad, Serbia. They investigated the manufacture

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of milk-based beverages obtained by Kombucha application. Local Kombucha culture was grown up on three substrates: sweetened black and green tea. Their concentrates were obtained by vacuum-evaporation and amounts of 10% and 15% (v/v) were applied to milk (2.2% fat). The traditional yoghurt starter (B3) was applied for producing control samples. All fermentations were stopped when the pH reached 4.4. Fermentation curves were registered, linear for yoghurt and sigmoidal for Kombucha. Two times faster process was achieved with yoghurt starter. Influence of inoculums concentration on the rate of fermentation was insignificant. Viscosities were higher for Kombucha beverages at lower speeds of spindle, but lower at higher speeds of spindle. Very high sensory scores were achieved for all beverages, after production and after 5-days’ storage.(Rodomir V. Malbasa, 2009)

Comparative Analysis of Indian Paneer and Cheese Whey for Electrolyte Whey Drink (Nupur Goyal and D.N.Gandhi 2011) had done in Dairy Microbiology Division, National Dairy Research Institute, Karnal, India. The present study was undertaken to make crude comparison between whey obtained from paneer and cheese during manufacturing. Paneer and cheese whey were compared in terms of all the minerals as well as physiochemical properties indispensable for electrolyte drink. The slight differences attributed, among various parameters can be mainly due the difference in manufacturing process. Our results indicated significantly higher concentration of sodium, potassium, calcium and chloride contents in paneer whey than cheese whey. The analysis is important as paneer whey can be utilized more efficiently otherwise creating environmental pollution especially in India.

Biology Department, Federal University of Lavras, 37200-000 Lavras, MG, Brazil has done the Production of fermented cheese whey-based beverage using kefir grains as starter culture: Evaluation of

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morphological and microbial variations.(Karina Teixeira Magalhaes ,2010) Whey valorization concerns have led to recent interest on the production of whey beverage simulating kefir. In this study, the structure and microbiota of Brazilian kefir grains and beverages obtained from milk and whole /deproteinised whey was characterized using microscopy and molecular techniques. The aim was to evaluate its stability and possible shift of probiotic bacteria to the beverages. Fluorescence staining in combination with Confocal Laser Scanning Microscopy showed distribution of yeasts in macro-clusters among the grain’s matrix essentially composed of polysaccharides (kefiran) and bacteria. Denaturing gradient gel electrophoresis displayed communities included yeast affiliated to Kluyveromyces marxianus, Saccharomyces cerevisiae, Kazachatania unispora, bacteria affiliated to Lactobacillus kefiranofaciens subsp. Kefirgranum, Lactobacillus kefiranofaciens subsp. Kefiranofaciens and an uncultured bacterium also related to the genus Lactobacillus. A steady structure and dominant microbiota, including probiotic bacteria, was detected in the analyzed kefir beverages and grains. This robustness is determinant for future implementation of whey-based kefir beverages.

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4. IN GENERAL, INGREDIENTS OF WHEY BEVERAGES4.1. Whey protein ingredient considerations

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Ingredients of whey

beverage


The most important component of an acidified whey protein RTD (Ready To Drink) beverage is obviously the whey protein ingredient .The key factors to be considered when selecting a whey protein are a) the whey protein’s method of isolation which determines the composition of the WPC (Whey Protein Concentrate or the WPI(Whey Protein Isolate) and b) a consistent source and manufacturing process to produce the ingredient. The compositional differences between ion exchange and membrane filtered WPI are explained in other publications available from USDEC, ingredients suppliers and other sources. Primary differences are the mineral and the glycomacropeptide content, both of which may affect suitability for a particular application. Whey protein concentrates 80% (WPC 80) are manufactured by membrane filtration processes. Fat and ash contents can vary among WPC 80s, as can flavor profiles. From a nutritional standpoint, a manufacturer will want to select the ingredient to which best matches their requirements: from total protein or mineral concentration to the presence of a particular whey fraction or amino acid. The best strategy for a manufacturer is to work closely with suppliers at the very early stages of the development process. Many U.S. suppliers offer guidance, typical formulations and technical assistance to support their customers when developing products. It is important to strive for a consistent lot-to-lot ingredient supply, and it may be necessary to develop a simple test which describes performance relative to the intended use, which goes beyond information provided in a standard specification or certificate of analysis. This is particularly true if the product and process are less robust to variations and if the beverage contains a protein level on the high end of the practical range. Again, close and early collaboration with a U.S. supplier is an important success factor.

4.2. Non-protein ingredient considerations

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Following are some of the other categories of ingredients frequently required or desired in whey protein RTD beverages. Their careful selection and laboratory evaluation are important when developing a shelf-stable product with excellent flavor and good consumer appeal. In all cases, please check country specific regulations when formulating these products to ensure. Compliance with all local requirements

1. Acidulants

Whey protein’s strong buffering capacity requires the use of considerable amounts of acid in the formula to bring the starting pH from around 6.5 down to 3.5 or lower. The most common acids used for making high-acid whey protein beverages are:

1) Phosphoric – a strong acid with a fairly plain flavor impact.

2) Hydrochloric – a strong acid with less desirable palatability, but may be used in medical nutritionals because it is the same acid found in the gastric system.

3) Citric – a weaker acidulant but very desirable for its contribution to the overall flavor profile of a fruit-flavored beverage. Citric acid is not recommended as the sole acidulant for very high protein drinks because of the extreme tartness imparted when used at high levels.

4) Malic – a weaker acid similar to citric acid but useful as an adjunct to formulas with apple or berry flavors, due to its natural presence in those fruits.

2. Carbon Dioxide (carbonation)

Carbonation is featured in this section because it should indeed be considered an ingredient, as much as a process, and because of its impact on acidity. There is increasing interest in improving the nutritional profile of carbonated soft drinks by adding whey protein.

3. Flavors

Whey proteins, unlike some vegetable protein sources, are widely compatible with, and even complementary to, many popular flavors. Acidified whey protein beverages are also less prone to the flavor adsorptive effects of other proteins in beverages, a phenomenon which would require heavier flavor usage (and cost).

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4. Sweeteners

There are many choices of caloric and natural or artificial reduced-calorie and non-caloric sweeteners suitable for use in whey protein beverages.

These include:

1) Sugars such as sucrose, fructose and high fructose corn syrup.

2) Sugar alcohols such as lactate and erythritol.

3) Artificial high-intensity sweeteners including sucralose and acesulfame potassium.

4) Natural high-intensity sweeteners such as those derived from citrus extracts.

The selection of sweeteners can impact mouth feel and protein stability in a formula specific manner. However, the choice of a sweetener is usually directed by calorie and flavor requirements. Note that in a particular protein RTD formulation, one sweetener may work well as the sole source of sweetness, but a combination of two often provides the best overall sweetness impact and compatibility with the base flavor.

6. Colors

Colors may be either artificial or natural, with light stability an important Consideration when using transparent or translucent bottles. The slowed gradation of ascorbic acid (Vitamin C) in beverages can, via its peroxide breakdown product, slowly decolorize beverages during shelf life. Color suppliers can offer guidance to manufacturers during the development process.

The name of colour which are used in whey beverage formulation:

Sr. No Name of colour EEC No. Availability1 Annatto E160B Liquid/ Powder2 Caratenaid E160A Liquid3 Marri Gold E161B Paste4 Beetroot/ Carrot E162 Liquid/ Powder5 Saff - Liquid/ Powder

7. Fruit Juices

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Juices are an excellent choice for creating flavorful whey protein isolate beverages and increasing consumer appeal. The use of natural juices may affect pasteurization requirements. The pH of the whey protein isolate in solution should be adjusted with the appropriate acidulant systems before combination with juices, because the protein will otherwise buffer the juice acids and possibly irreversibly change the product characteristics.

8. Minerals

The stability and clarity of acidified whey protein beverages is believed to be affected by the amount of mineral ions, such as sodium or calcium, present in the system. Therefore, mineral selection and level of fortification may be limited by their effect on the final beverage. In general, adding salts increases aggregation in thermally processed whey beverages, thus decreasing stability.

8. Vitamins

As with any food or beverage product, vitamin must be chosen and formulated according to their compatibility with the overall system .Most water-soluble vitamins are fairly stable in acid environments. However, consideration must be given to color and flavor contribution, processing losses and light stability for a RTD in a transparent or translucent bottle. Ingredient interactions should also be considered. Generally vitamin B1, Vitamin B2, Vitamin C etc.

9. Stabilizers and Emulsifiers

Stabilizers and emulsifiers can be very important to neutral, shake-type beverages, especially when mixtures of proteins and/or cocoa powder are used. Carrageenan, cellulose gel and cellulose gum are stabilizers used in neutral Beverages with added protein. Pectin is used for whey protein beverages in the pH range between 3.5 and 4.6 to protect and stabilize the proteins during thermal processing and throughout their shelf life. Stabilizers are generally not needed below pH 3.5 in acidified whey protein isolate RTD beverages. Emulsifiers like mono- and diglycerides and buffers such as tetrasodium pyrophosphate are commonly used in neutral pH beverages using whey proteins along with other milk proteins. Establishing the ideal levels of stabilizers, buffers and emulsifiers are especially important to ensure long-term stability of protein fortified beverages in the acid and neutral category.

9. Preservatives

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Acidified whey protein beverage formulas can include chemical preservatives such as sorbets and benzoates to control the growth of yeasts, molds and bacteria that could lead to product spoilage.

10. Nutraceuticals

Whey protein drinks are considered high-value nutritional beverages, and are often fortified with additional nutritional components such as plant sterols to lower cholesterol, lutein to improve eye health, orma-huang or guarana which are reported to boost energy. Live and active cultures are frequently incorporated into cultured dairy beverages containing whey proteins. This latter category is usually pasteurized, cultured and stored refrigerated to preserve the probiotic health effects, although some products do receive heat treatment and are thus shelf-stable. (Steve Rittmanic, 2006)

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Beverage Formulatio

n


5. BEVERAGES FORMULATIONS

The formulations in this section are provided as a starting point for product development purposes. Adjustments may be necessary, depending upon the exact nature of ingredients used, processing and storage variables, local regulations, and target consumer preferences in each market. The formulations are courtesy of the Dairy Ingredients Applications Laboratory, Wisconsin Center for Dairy Research, Madison, Wisconsin, USA. The laboratory is supported by Dairy Management Inc., Rosemont, Illinois, USA and the Wisconsin Milk Marketing Board. (Steve Rittmanic, 2006)

1. Isotonic Drink with WPI (Whey Protein Isolate):

Ingredients Usage Level (%)Water 85.43Fructose 9.00WPI 5.00Phosphoric acid 0.37Natural mango flavor 0.05Yellow color 0.04Potassium sorbate 0.04Salt 0.04Calcium Chloride 0.02Potassium Chloride 0.01Total 100

Procedure:

1. Reconstitute WPI in formula water (at ambient temperature) with a high speed mixer and allow hydrating 20minuteswith little agitation.

2. Mix in fructose, salts, flavor and color.

3. Use 85%solution of acid to adjust pH to 3.2.

4. Heat to 90ºC (195°F) for 45 seconds.

5. Fill containers and cool to 4ºC (40°F).

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Nutritional Content per 100 grams

Calories 50 kcalTotal Fat 0 g Saturated Fat 0 g Trans Fat 0 gCholesterol 0 mgSodium 20 mgTotal Carbohydrate 9 g Dietary Fiber 0 g Sugars 9 gProtein 5 g Vitamin C 0 g Vitamin B1 0 g Vitamin B2 0 g Calcium 2 g

2. Low pH Juice Drink with WPI (Whey Protein Isolated)

Ingredients Usage Level %Water 80.73High Fructose corn Syrup 9.40WIP 4.70Apple Juice Concentrate -70 Brix 4.70Phosphoric Acid Solution-85% 0.35Natural Berry Flour 0.10Red Color 0.02Total 100

Procedure:

1. Reconstitute WPI in formula water (at ambient temperature)with a high speed mixer and allow to hydrate for 20minutes.

2. Mix in high fructose corn syrup, juice, flavor and color.

3. Use 85%solution of acid to adjust pH to 3.2.

4. Heat to 90ºC (195°F) for 45 seconds.


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Calories 60 kcalTotal Fat 0 g Saturated Fat 0 g Trans Fat 0 gCholesterol 0 mgSodium 0 mgTotal Carbohydrate 11 g Dietary Fiber 0 g Sugars 7 gProtein 4 g Vitamin C 0 mg Vitamin B1 0 mg Vitamin B2 0 mg Calcium 0 mg

3. Meal Replacement Drink with WPC 80(Whey Protein Concentrate)

Ingredients Usage Level(%)Skim milk 93.00Granulated Sugar 4.70WPC 80 1.40Vanilla Extract 0.50Mono & Diglycerides 0.20Carrageenan 0.10Tetrasodium pyrophosphate 0.10Total 100

Procedure:

1. Disperse all ingredients into skim milk at 4ºC (40°F) with a high-speed mixer.

2. Check pH and adjust to 7.0-7.1 by adding tetra sodium pyrophosphate.

3. Hydrate for 20minutes.

4. Check pH and re-adjust to 7.0-7.1 if necessary by adding tetrasodium pyrophosphate.

5. Heat to 85ºC (185°F).

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6. Homogenize: first stage at 250 Bar (24.82MPa, 250kg/cm2 or 3600psi) and second stage at 48 Bar (4.82MPa, 49kg/cm2 or 700psi).. Cool to 25ºC (77°F).

7. Bottle.

8. Retort with rotation at 10rpmat120ºC (250°F) for 4 to 5minutes.


Calories 60 kcalTotal Fat 1 gSaturated Fat 0.5 gTrans Fat 0 gCholesterol 5 gSodium 110 mgTotal Carbohydrate 9 gDietary Fiber 0gSugars 9 gProtein 4 gVitamin C 0 gVitamin B1 0 gVitamin B2 0 gCalcium 120 g

4. Low Sugar Drink with WPC 80

Ingredients Usage Level (%)Water 90.62WPC 80 5.29Cream 2.11Pectin 1.37Phosphoric acid 0.28Mango Flour 0.20Sucralose 0.10Red Colour 0.02Yellow Colour 0.01Total 100

Procedure:

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1. Hydrate stabilizer in half of the formula water at 85ºC (185°F) and let swell for 10minutes.

2. Agitate at 85ºC (185°F) until completely dissolved; allow cooling to 60ºC (140°F).

3. At the same time, reconstitute WPC in the remaining formula water at ambient temperature with a high-speed mixer, add cream and let hydrate for 20minuteswith little agitation.

4. Add WPC solution to stabilizer solution and add sweetener, flavor and colors.

5. Use 85%solution of acid to adjust pH to 3.8.6. Homogenize: first stage at 250 bar (24.82MPa, 250kg/cm2 or 3600psi)

and second stage at 48 bar (4.82MPa, 49kg/cm2 or 700psi).7. Heat to 88ºC (190°F) for 45 seconds. Cool to 24ºC (75°F).8. Fill containers and cool to 4ºC (40°F).

Nutritional Content per 100 grams:

Calories 30gTotal Fat 1gSaturated Fat 0.5gTrans Fat 0gCholesterol 5gSodium 15gTotal Carbohydrate 2gDietary Fiber 0gSugars 0gProtein 4gVitamin C 3.6gVitamin B1 0.02gVitamin B2 0.05gCalcium 24g

5. Juice Drink with WPC 80(Whey Protein Concentrate)

Ingredient Usage Level (%)

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Water 79.57Granulated Sugar 8.33WPC 80 5.20Juice concentrate ,Pectin 5.09Pectin 1.30Milk Calcium 0.31Phosphoric acid 0.20Total 100

Procedure:

1. Hydrate stabilizer and sugar in half of the formula water at 85ºC (185°F) and let swell for10minutes.

2. Agitate at 85ºC (185°F) until completely dissolved; allow cooling to 60ºC (140°F).

3. At the same time, reconstitute WPC and milk calcium in remaining formula water at ambient temperature with a high-speed mixer and let hydrate for 20 minutes with little agitation.

4. Add juice, WPC and milk calcium solution to stabilizer solution. 5. Use 85%solution of acid to adjust pH to 3.8.

6. Homogenize: first stage at 250 bar (24.82MPa, 250kg/cm2 or 3600psi) and second stage at 48 bar (4.82MPa, 49kg/cm2 or 700psi).

7. Heat to 80º (175°F) for 45 seconds. Cool to 24ºC (75°F).

8. Flavor with juice concentrate and add colors for desired tint.



Calories 70g

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Total Fat 0g Saturated Fat 0g Trans Fat 0gCholesterol 0gSodium 20gTotal Carbohydrate 12g Dietary Fiber 0g Sugars 11gProtein 4g Vitamin C 9g Vitamin B1 0.02g Vitamin B2 0.05g Calcium 96g

(Steve Rittmanic, 2006)

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Material and

Methods


6. MATERIALS AND METHODS

6.1. Materials

1. Sweet Liquid Whey (70%)

2. Whey Protein Concentrates 35% (w/w)

3. Sugar 15% and Colour 0.05%

4. Mango Juice Concentrate (30%)

5. Essences (0.5%)

6. Anti-Foaming Agent (0.5%)

7. Packaging Material i.e. Plastic Bottles

6.2. Chemicals

1. Lactic Acid 2% (w/v)2. Calcium Lactate (For pH Regulation)

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6.3. Whey Processing:

Fig.6.1. Flow chart of whey processing (Ref. Mirjan Djuric 2004)

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Recieving of Whey

Stored temporarily at about 500C

Casein fines recovary & fat separation

Pasteurisation at 850C For 10min.

Sterilization 90-920CFor 5min

Homogenization

Filtration & Clarification

Cooling at 10-150C

Cold Storage

By Ultrafiltration

Fat Content 25-30%


The process for manufacturing whey beverage consists of the following steps:(Singh S. Ladkani, 1994)

1. Collection of whey and its standardization :The whey obtained from cheese and paneer making is passed through the cream separator to remove fat and then heated to the appropriated temperature, cooled and is fed to incubation tank pre-adjusted to the specified temperature.

2. Culture preparation:Whey is sterilized by heating for specified time followed by cooling and inoculated with required amount of pure culture of the required species grown in litmus milk. It is further inoculated for the preparation of intermediate and bulk culture in the same manner.

3. Fermentation process:The cold heat treated whey is inoculated with a pure and active culture at a desired level. After inoculation when the acidity of the whey reaches at the desired level the fermented whey is cooled and filtered through the filter press.

4. Fortification with sugar and flavor:Filtered fermented whey is first fortified with sugar in the form clear sugar syrup and then flavored with the combination of pineapple and orange essence at the required level. No colour is added in the product as the colures are not stable in the product due to low pH of the finished product.

5. Packaging and storage:The prepared whey beverage is cooled and then it is filled in polypacks or glass bottles which are crown corked after filling, if intended for immediate consumption. For increased shelf life of product, beverage should be pasteurized before packaging or alternatively pasteurized in the container.

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Whey Beverage Manufacturing (In general):

Fig.6.2. Flow Chart of Beverage Manufacturing

(Ref. Article on Acido whey, 2010)

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Collection of whey

Standardisation

Sterilisation and Pasteurisation at 850C for 10min

Cooling at (15-200C)

Blending With Fruit Juice

Fortification With Sugar and Essence

Filtration And Clarification

Flavoring( Adjust PH 3.6-4.2 )

Bottling and Corcking

Packaging

Cold Storage


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Formulation of functional fermented whey based beverage using lactic acid bacteria


7. FORMULATION OF FUNCTIONAL FERMENTED WHEY-BASED BEVERAGE USING LACTIC ACID BACTERIA(Case Study)

7.1. Preparation of media:

1. Microorganisms and media

The strains L. acidophilus CRL 636, L. delbrueckii subsp. Bulgaricus CRL 656 and Streptococcus thermophilus CRL 804 used in this work were obtained from the Culture Collection of Centro de Referencia para Lactobacillus (CERELA), San Miguel de Tucumán, Argentina. Cultures were stored at −20 °C in 10% (w/v) sterile reconstituted skim milk containing 0.5% (w/v) yeast extract, 1.0% (w/v) glucose and 10% (v/v) glycerol. Whey protein concentrate 35%, w/w protein (WPC35), powder (kindly provided by MILKAUT S.A., Argentina) was reconstituted with distilled water to 10% (w/v) and the pH was adjusted to 8.0 with 2 mol/l NaOH. The reconstituted WPC35 was heat treated at 116 °C for 20 min, stored at 4 °C until use (no longer than one week) and

used as fermentation medium.

The presence of deteriorating microorganisms was assessed by plating pure or diluted (ten times) beverage samples in Baird Parker agar supplemented with egg yolk and tellurite (for Staphylococcus aureus), violet red bile lactose agar (VRBA, for total coliforms), plate count agar (PCA, for mesophilic microorganisms), and potato dextrose agar (PDA, for fungi and yeasts). All media were purchased from Britania S.A (Buenos Aires, Argentina). Plates were incubated according to the manufacturer's indications.

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2. Fermentation conditions

Cultures were transferred twice in WPC35 prior to experimental use; 16 h old cultures (2% v/v) were used as inoculums individually, or combined as follows: L. delbrueckii subsp. bulgaricus CRL 656: S. thermophilus CRL 804: L. acidophilus CRL 636 at a 1:1.5:6.4 CFU/ml ratio(Panesar,2007). Fermentations were performed statically in sealed bottles containing 300 ml of WPC35 and incubated at 37 °C for 24 h.Samples were aseptically withdrawn every 2 h during 12 h and at 24 h of incubation. Cell viability was determined by plating appropriate dilutions of the cultures in MRS agar (MRS Britania, Buenos Aires, Argentina, plus 15 g/l agar). To determine the viable cell count of the L. acidophilus strain in the mixed culture, 1.5% (w/v) bile salt (Sigma Chemical CO, St. Louis, USA) was added to MRS agar (Vinderola and Reinheimer, 2000). The strains L. delbrueckii subsp. bulgaricus CRL 656 and S. thermophilus CRL 804 were selectively counted by means of their shape in the mixed culture by plating the fermented WPC35 in MRS agar (aerobic conditions) as recommended by the International Dairy Federation for the selective count of L. delbrueckii subsp. bulgaricus and S. thermophilus in yogurt. (Vinderola and Reinheimer 1999). Plates were incubated at 37 °C for 48 h and colony-forming units (CFU)/ml were determined. Cells of L. delbrueckii subsp. bulgaricus CRL 656 appeared as irregular white large colonies while those of S. themophilus CRL 804 as small round white colonies. To confirm the identity of the colonies, cell morphology was observed with an Olympus Vanex microscope (Tokyo, Japan). Cell count was expressed as log CFU/ml. Decrease in pH was followed with a digital pH meter (Altronix TPX 1) every 2 h during the first 12 h and after 24 h incubation.

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7.2. Fermented Whey Beverage Formulation:

1. WPC35 was allowed to ferment for 12 h.

2. Cooled down in ice and diluted 1:3 with peach juice.

3. Previously dissolved in sterile water or calcium lactate 2% (w/v).

4. Calcium lactate was added as acidity regulator following the indications of the Codex Alimentarius (CODEX STAN, 192-1995).

5. The resulting beverages were distributed in sterile plastic bottles in triplicates and stored at 10 °C for 28 days.

6. Viable cell count, pH, sugar and lactic acid concentrations, proteolytic activity, free amino acid content and whey protein degradation were determined after 0, 7, 14, 21 and 28 days of storage.

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WHEY PROTEIN CONCENTRATE (35%)

RECEIVING OF MANGO FRUIT


Fig.7.1. Flow chart of Functional whey beverage formulation

(Ref. Micaela Pescuma, 2010)

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RECONSTITUTE WPC WITH DISTILLED WATER BY 10%(W/V)

TRANSFERRED INTO CULTURE

INCUBETED AT 370C FOR 24hr

FERMENTATION

BOTTLING OF WHEY

SHORTENING OF FRUIT

REMOVING OF CROWN

CUTTING INTO SMALL PIECES

PULPING IN MIXER

PASSING INTO HAND PULPER

PASSING INTO HAND PULPER

BLENDING OF WHEY AND MONGO JUICE (70:30)

DISSOLVED 2% CALCIUM LACTED (w/v)

FORTIFICATION WITH SUGAR(PH 4.2-5)

FILTRATION AND CLARIFICATION

ASEPTIC PACKAGING & STORAGE(8-100C)

BOTTLING AND CARKING


8. QUALITY CONTROL AND TESTING

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Quality Control and

Testing


1. Analysis of metabolites

Sugar content (lactose, galactose and glucose) and organic acids (lactic, acetic, and formic) production were analyzed during fermentation by High Performance Liquid Chromatography (HPLC). HPLC was performed using a Knauer Smartline System HPLC (Berlin, Germany) with a Knauer Smartline RI detector fitted with a Biorad Aminex HPX-87H column (300×7,8 mm, Hercules, CA, USA). The operating conditions were the following: 5 mol/l H2SO4 was used as fluent at a flow rate of 0.6 ml/min during 30 min and an internal temperature of 45 °C. For the quantification of sugars and organic acids, calibration curves for each compound were performed using pure standards at different concentrations.

2. Proteolysis assessment

The proteolytic activity of LAB was measured in samples of fermented WPC35 (every 2 h during 12 h and at 24 h) and of the beverage during storage (0 and 28 days) by using the o-phthaldialdehyde (OPA) test (Church et al., 1983). The increase in optical density at 340 nm (OD340) relative to the control was determined using a VERSA max Tunable Microplate reader (Sunnyvale, CA, USA). The OPA solution contained: 2.5 ml of 20% (w/v) SDS, 25 ml of 100 mole/l sodium tetraborate (Sigma Chemical Co), 40 mg of OPA (Sigma Chemical Co) (previously dissolved in 1 ml methanol), 100 μl of 2-mercaptoethanol (Merck, Darmstadt, Germany) and distilled water up to a 50 ml final volume. Fermented samples were incubated with 0.75 mol/l trichloroacetic acid (Sigma Chemical Co) at a sample: trichloroacetic acid ratio=1:3 at 4 °C for 30 min and centrifuged (5000 rpm 10 min). Ten microliters of the supernatant of this mixture was added to 0.2 ml of OPA reagent and then incubated at room temperature for 5 min until the OD340 was read in the microplate spectrophotometer. Proteolytic activity was arbitrarily expressed as μg leucine (Leu) released/ml using a standard curve of L-leucine (BDH Chemicals Ltd Poole, England).

3. Free amino acid determination

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The free amino acid content of non-fermented and fermented WPC35 as well as the stored beverage was determined. Samples were treated to eliminate proteins and the amino acids were extracted as described by Jones et al. (1981). The reaction was prepared by mixing 200 μl of WPC35 with 2% (w/v) of SDS (dissolved in 0.4 mol/l sodium borate buffer, pH 9.5) and 200 μl of the OPA methalonic solution. The mixture was shaken, incubated for 1 min and the reaction was stopped by adding 400 μl of 0.1 mol/l sodium phosphate buffer (pH 4.0) and filtered through 0.2 μm nylon membrane (All tech Associates Inc., Deerfield, IL, USA). The amino acids used as standards (Sigma Chemical Co) were treated in the same way as the above samples. The amino acid content of the samples was analyzed by reverse phase-high performance liquid chromatography (RP-HPLC) with an ISCO model 2360 (ISCO, Inc., Lincoln, NE,USA) fitted with an Ultrasphere ODS C18 column (4.6×25mm, particle size 5 μm, Beckman Instruments Inc., Fullerton, CA, USA). The equipment was coupled with an ISCO model 2350 pump and an ISCO FL-2 fluorescence detector (ISCO Inc.). The operating conditions were the following: flow rate, 1.7 ml/min; solvent A, tetrahydrofurane: methanol: sodium acetate (1:19:80, v/v/v) 0.05 mol/l pH 5.9 (Sigma Chemical Co.) in ultra pure water; solvent B, methanol: sodium acetate 0.05 mol/l pH 5.9 (80:20 v/v) (Sigma Chemical Co). Elution was performed by applying a linear gradient of 100% solvent A over 1 min, then 0–50% solvent B over the following 20 min, and 50–100% solvent B over the last 20min. Absorbance was recorded at 305–395 and 430–460 nm excitation and emission wavelengths, respectively. The injection volume of derivative amino acids was 10 μl. The HPLC was coupled with the software Chem. Research 150 Data System 3.0.2. All the amino acids, except proline, cystein and methionine, were determined under the assayed conditions. Amino acid concentration was expressed in μg/ml.

4. Hydrolysis of β-lactoglobulin in WPC35

Degradation of β-lactoglobulin was monitored by RP-HPLC using a Knauer Smartline System (Manager 5000, pump 1000) with a UV detector (2000) fitted with a C18 column (Pursuit 4.6×250 mm, 300 A, 5 μm, Varian, Lexington, USA). The method used included buffer A: water/acetonitrile/trifluoroacetic acid (90/10/0.1, v/v/v), and buffer B acetonitrile/trifluoroacetic acid (100/0.1, v/v) with a flow rate of 1 ml/min. The gradient used was 100% buffer A up to 10 min and 10 to 60% buffer B in a linear fashion between 10 and 60 min. Eluted peaks in the chromatograms were detected at 214 nm. Samples for

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HPLC were prepared as follows: WPC35 samples (fermented and nonfermented) were mixed 1:1 with reduction buffer containing urea and 20 mole/l dithiothreitol (DTT) and incubated for 60 min at 30 °C. Prior to injection in the column, the reduced sample was diluted 5-fold in buffer a containing 6.0 mol/l urea. BLG hydrolysis was expressed as percentage and was calculated by measuring its relative peak area with respect to the control (non-fermented sample). 2.8. Strains compatibility Strains compatibility was evaluated by the plate diffusion assay (Parente and Zottola, 1991). Briefly, overnight cultures grown in MRS were washed twice with saline solution and suspended at the initial volume. Plates were prepared by pouring 15 ml of MRS soft agar (MRS plus 0.7%, w/v, agar) containing 60 μl of the cell suspension on the agar. After overlay solidification, 5 mm diameter wells made with sterilized plastic straws were inoculated with 60 μl of culture supernatants from the other strains. After incubation at 37 °C for 16 h, appearance of inhibition zones were observed.

5. Statistical analysis

All assays were carried out in triplicate, and results were expressed as mean values with standard deviations. Statistical analyses were performed using MINITAB 14 software. Comparisons were accomplished by ANOVA general linear model followed by Turkey’s post-hoc test, and pb0.05 was considered significant.

6. pH

The pH values were measured using a Fisher Accument Model AP 63 pH meter with a pH/Automatic Temperature Compensation (ATC) combination electrode calibrated at pH 7.0. Measurements of pH were taken on the uncarbonated and carbonated samples in duplicates and an average was calculated.

7. Brix

Soluble solids content was measured using an Abbe Mark2 Refractometer. Samples were loaded into the machine and viewed through the eyepiece for adjustment until the color zone separation matched the cross-hair line. The % Brix was then read directly from the display and recorded. Duplicate measurements were taken from both the uncarbonated and carbonated samples and then averaged.

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8. Color

Color measurements were made using a HunterLab MiniScanMS/S-4000S spectrocolorimeter (Hunter Associates Laboratory, Inc., Reston, VA) to determine the L*, a*, and b* values of the uncarbonated and carbonated samples. The MiniScan™ was standardized with a black light trap and white color tile before the first sample was measured. Samples were placed in a polysterene clear cup with a plastic ring inside holding the sample in place and then covered with a black cup to prevent light from reflecting and interfering with the measurement. L*, a*, and b* values were measured three times and then averaged. Duplicates were made with all samples. L* represents the whiteness of a sample, where 100 represents white and 0 is black. A positive ‘a*’ value indicates redness, and a negative ‘a*’ value indicates greenness. A positive ‘b*’ value represents yellowness and a negative ‘b*’ value represents blueness.

9. Fat Determination

Apparatus:

Soxhlet Apparatus with a 250 ml. flat bottom flask.

Reagents:

(a) Petroleum Ether – Boiling point 40 to 80 ºC

(b) Benzene- Alcohol–Phenolphthalein Stock solution – To One liter of distilled benzene add one liter of alcohol or rectified sprit and 0.4 gm of phenolphthalein. Mix the contents well.

(c) Standard potassium hydroxide solution – 0.05 N.

Weigh accurately sufficient amount. of biscuit powder ( 20 – 25 gm) which will yield 3- 4 gm of fat and transfer it to a thimble and plug it from the top with extracted cotton and filter paper. In case of filled and coated biscuits the weight of the biscuits includes the filling and coating material. Dry the thimble with the contents for 15 to 30 minutes at 100 ºC in an oven. Take the weight of empty dry Soxhlet flask. Extract the fat in the Soxhlet apparatus for 3 to 4 hours and evaporate off the solvent in the flask on a water bath. Remove the traces of the

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Procedure:


residual solvent by keeping the flask in the hot air oven for about half an hour. Cool the flask. Weigh accurately about 3.0 gm of extracted fat in a 250 ml conical flask and add 50 ml. of mixed benzene-alcohol-phenolphthalein reagent and titrate the contents to a distinct pink color with the potassium hydroxide solution taken in a 10 ml. micro burette. If the contents of the flask became cloudy, during titration add another 50 ml of the reagent and continue titration. Make a blank titration with 50 ml. of the reagent. Subtract from the titer of the fat, the blank titer. (Ranganna, S.1986)

10.Carbonation Level

Volumes of carbon dioxide in the carbonated beverage were measured using a Series 6000 Zahm Model D.T. Piercing Device (Zahm and Nagel, Holland, NY). The full bottle of carbonated beverage was inverted and subsequently pierced with the device. The pressure inside the bottle was measured directly from the accompanying gauge. The device’s thermometer was inserted into the product and the temperature was read directly. Based on a table provided with the carbonation equipment, the total volume of carbon dioxide dissolved in the product was calculated. Duplicate measurements were taken from the carbonated sample and then averaged.

11.Microbiological

AOAC methods 986.33 and 989.10 for dairy products were used to detect the presence of aerobic bacteria in both the uncarbonated and the carbonated beverage products. One milliliter of the product, diluted to 1:10 and 1:100 with serial dilutions, was plated on Petrifilm (Aerobic Count Plates, 3M, and MN) and then incubated at 32°C ± 1°C for 48h ± 3h. AOAC 997.02 method was used to detect the presence of yeast and mold in both the uncarbonated and carbonated beverage products. One milliliter of the product, using the previously stated serial dilutions, was plated using 3M™ Petrifilm (Yeast and Mold Plates, 3M, MN) and incubated at 20-25°C for 3 to 5 days. AOAC method 983.25 was used to detect the presence of total coli forms in both the uncarbonated and carbonated beverage products. One milliliter of the product, using the previously stated serial dilutions, was plated on 3M Petrifil (ECC Plates, 3M, and MN) and then incubated at 32°C ± 1°C for 48h ± 3h.

12.Sensory

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Faculty, staff and students participated in an informal sensory study. Consumers (n=35) were asked to fill out a questionnaire regarding age, gender, and beverage consumption habits. An informed consent form listing ingredients and potential ingredients in the beverage was signed by each participant prior to tasting. Consumers were asked to evaluate the sample for overall liking, appearance, mouth feel, flavor, and sweetness on a 9-point hedonic scale anchored on the left by 1 (“dislike extremely”) and on the right by 9 (“like extremely”).

13.Nutrition Labeling

The basic nutritional content was determined for the carbonated beverage using the Genesis R&D labeling program (ESHA Research, Salem). The serving size was reported by FDA.

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9. Machineries1. Fermenter

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Machineries


2. Centrifuge for separation of fat3. Pasteurizer4. Filling Machine5. Cold Storage-Refrigerator

1. Fermenter:

(Ref.Athor,Dr.J.N.de Wit, 2009)

Fermenters that is procured from reliable manufacturers. The range of Fermenters offered by us is renowned for its sturdy construction and durability. Further, the range of Fermenters offered by us does not react with chemicals, thereby offering high utility and effectiveness to the clients. Our range of Fermenters is available in various capacities to ensure high utility to the clients. The high quality and effective performance of these Fermenters have made them the preferred choice across industries globally.

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2. Centrifuge :


The Centrifuges that is widely used to separate substances of different densities. The Centrifuges offered by us have been fabricated using high grade raw material. Further, we follow an extensive quality management program to ensure compliance of our range with various international quality standards. The range of Centrifuges offered by us is renowned for its noiseless and effective performance, and is widely used in chemical as well as pharmaceutical industries. Since our range of Centrifuges has been manufactured as per international safety norms it offers enhanced safety.

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http://www.google.co.in/imgres?imgurl=http://www.niroinc.com/images/food_chemical/spray_dryer_two_stage.jpg&imgrefurl=http://www.niroinc.com/food_chemical/spray_drying_foodstuffs.asp&usg=__hESutLYSAKn5wYHsckGqB5vocWM=&h=250&w=230&sz=10&hl=en&start=10&zoom=1&tbnid=udT6qK5yXnCyfM:&tbnh=111&tbnw=102&ei=p7CiTYeBKJGKvgPS_NmLBQ&prev=/search?q=spray+dryer&um=1&hl=en&biw=1345&bih=556&tbm=isch&um=1&itbs=1


1. Pasteurizer


Pasteurizer equipment is used for pasteurization of whey and fruit juice. Pasteurization is done at 80-850C.

2. Filling Machine

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The bottle filling machine is capable to fill 60 /120 machines per minutes. The pump speed of these bottle filling machines is 60 / 120 strokes / min while filling speed depends on filled volume / bulk density of material.

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Material balance


10. Material Balance

Basis: 500ml of whey

F= W + P (1)

1. Sample Estimation:

Take 500ml of whey as a feed.

There are number of process carried out on the raw whey such as Standardization, sterilization and pasteurization.

This process results into 10% loss of whey.

2. Fruit Juice:

30% (w/w) of fruit juice is used.

Therefore 30% of 500ml =500*(30/100) =150L.

The following are the process carried on fruit juice such as clarification, filtration etc.

3. Sugar:

15 %( w /w) sugars required.

Therefore 15% of (135+450) L = 87.75Kg of sugar

Addition of (1) + (2) + (3)

450+135+87.75= 672.75

4. Colour and Essences are added to 0.5%(w/w) each

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500L of wheyLoss Occurs (Standardization,

Pasteurization, sterilization)

0

450L whey remains

150L of whey Loss Occurs (Clarification, Filtration)

135L Fruit Juice remains


Therefore 672.75 X (0.5+0.5)/100 =6.7272 g

Therefore the net beverage production = 672.75+6.7272= 679.47L

After the preparation of whey beverage the operation such as bottle filling sterilization take place. Here 5% loss of whey beverage occurs.

Therefore the total whey beverage production 645.51L i.e. nearly 645L

Calculation for 1, 00,000 L Whey Beverage:

1. Whey:

Raw whey = 77519L

After 10% loss in processing

77519* (10/100) = 7751.9 L

Therefore, 77519-77519.9 = 69767.1 L of whey

= 69769L 0f whey (nearly) (2)

2. Fruit Juice:

Fruit Juice required 30 %( w/w)

77519*(30/100) = 23255.7 L

After some loss in fruit juice processing (10%loss)

After 10% loss in processing,

23255 – 2325.57 = 20930.13 L of fruit juice.

= 20930L of fruit juice (nearly) (3)

Addition of (1) & (2)

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Therefore, 20930 + 69767 = 90967L

3. Sugar:

Sugar required 15 %( w/w)

90967*(15/100) = 13645.05

= 13645L (in form of liq. Concentrate) (4)

Addition of (3) & (4)

Therefore, 90967 + 13645 = 104612L

4. Colour & Essence:

Flavour and Essence required 0.5 %( w/w)

Therefore, 104612 + 1046.12(Colour & Essence)

= 105658.12L

= 105658L (nearly)

5. Packaging & other process

After 5%loss in that process

105658*(5/100) = 5282.9L

Therefore, 105658 – 5282.9 = 100375.1L

= 1, 00,000L (nearly)

Material balance for the manufacturing of whey beverage is as follows:

Feed = Product + Waste

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Therefore,

(77519+23255.7+13645+1046.12) L = (100375+5090.82) L

105465.82L = (100375+5090.82) L

= 1, 00,000L (approx.)

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11. COST ESTIMATION

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Cost estimation


1. Fixed Capital:

A) Equipment Cost

Equipment

SN. Equipment Cost(Lakh)

1 Fermenter 50L2 Pasteurizer 15L3 Sterilizer 10L4 Mixing Tank 0.8L5 Storage Tank 8L6 Inoculums Tank 12L7 Laboratory Equipment 3L8 Generator 10L9 Labeling Machine 0.8L10 Centrifuge 10L

Total 1,16,70,000Rs.

A) Direct Cost

F1 = Purchase equipment cost = 0.15F2 = Instrumentation and control = 0.10F3 = Piping (installed) = 0.15F4 = Electrical (installed) = 0.15F5 = Building (Including services) = 0.20F6 = Utilities = 0.50F7 = Yield Improvement = 0.05F8 = Auxillary Building = 0.15

TOTAL = 1.45

Total Direct Cost = Total X Equipment Cost

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=1.45 X 1, 16, 70,000

=16, 92, 1500 Rs.

B) Indirect cost

F9 = Engineering & Supervision = 0.20

F10 = Construction expenses = 0.15

F11 = Contractor Fees = 0.10

F12 = Contingency = 0.15

Total = 0.60

Indirect Cost = 0.60 X 1, 16, 70,000

= 70, 02,000

Total Fixed Cost = Equipment Cost + Direct Cost + Indirect Cost

= 3, 55, 93,500Rs.

Working Capital = 15% of fixed cost

= 0.15 X 3, 55, 93,500

= 53, 39,025Rs.

C) Land Cost

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According to Amravati MIDC (Three Star Zone)

Cost per Square feet = 90Rs.

Cost of 40,000square feet = 36, 00,000Rs.

D) Total Investment

= Fixed Cost + Working Cost + Land Cost

= 3, 55, 93,500 + 53, 39,025 + 36, 00,000

= 4, 45, 32,525Rs.

E) Variable cost

a) Raw material cost:1. Whey = 5, 42,633Rs.2. Sugar = 27,290Rs. 3. Fruit juice = 6, 97,671Rs. (as per 30Rs./lit)4. Colour and Essence = 32,406Rs.

Total = 13, 00,000Rs.

b) Miscellaneous Material = 10% of Working capital = 5, 33,902.5Rs.

c) Utilities (Fresh Water, Electricity) = 10% of fixed cost = 35, 59,350Rs.

d) Transportation Cost = 5% of raw material = 65,000Rs.

Total Variable Cost = (a) + (b) + (c) + (d)

= 54, 58,252.5Rs.

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F) Other Fixed Cost

a) Maintenance 10% of fixed cost = 35, 59,350.0Rs.

b) Operating labour and staff = 52, 00,000Rs.c) Plant over head = 60% 0f fixed cost

= 2, 13, 56,100Rs.d) Insurance = 1% of foxed cost

= 3, 55,935Rs.e) Royalty = 2% of fixed cost

= 7, 11,870Rs.

Total = 2, 84, 83,255Rs.

Direct production cost = Variable cost + Other Fixed cost

= 54, 58,252.5 + 2, 84, 83,255

= 3, 39, 41,507.5Rs.

G) Sale Income

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Whey Beverage produced per year = 1, 00,000 L/ yr.Price of Whey Beverage = (5458252.5/1, 00,000) = 54.5 Rs/ L Therefore the cost of whey beverage per litre is 54.5Rs./lit.

Total Income in a year = 5, 45, 00,000 Rs.

F) Profitability Analysis

a) Gross Return = sale income – production cost

= 5, 45, 00,000- 3, 39, 41,507.5

= 2, 05, 58,492.5 Rs.

b) Depreciation Cost = 10% of investment

= 20, 55,849.5Rs.

c) Taxable Income = Gross return – depreciation cost

= 2, 05, 58,492.5 – 20, 55, 849

= 1, 85, 02,643.25Rs.

d) Tax on Profit = 50% of taxable income

= 92, 51,321.625Rs.

e) Net Profit = Gross return– Depreciation cost-Tax = 2, 05, 58,492.5-2, 05, 58,449.25-92, 51,321.1625 = 92, 51,321Rs.

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PLANT LAYOUT

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Plant layout


This layout can play an important part in determining construction and manufacturing cost, and thus must be planed carefully with attention being given to future problems that may arise. Proper layout in each case will include arrangement of processing areas, storage areas and handling areas in efficient coordination and with regard to such a factors as:

1. New site development or addition to previously developed site.2. Type and quality of products to be produced.3. Type of process and product control.4. Operational convenience and accessibility.5. Economic distribution of utilities and services.6. Type of building and building-codes requirements.7. Health and safety considerations.8. Auxillary equipment.9. Space available and space require.10.Roads and railroads.

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14. Conclusion

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Conclusion


Whey is a liquid product separated out from milk in the preparation of paneer and cheese. The whey contains 6 to 7% of total solids comprising of approximately 70% of lactose, 0.9% of protein and trace amount of water soluble vitamins, minerals and fat. So far the whey is considered to be a waste product in the dairy industry but process has been developed to produce a healthy drink from this waste material. This beverage unlike the other carbonated beverages which are of little usefulness.

It has following advantages:i) It has a good nutritional value

ii) It has therapeutic values namely a. Protection against gastro-intestinal disorders b. Bio- availability of vitamins.

iii) It has three weeks shelf life under refrigeration.

iv) It is much cheaper in cost compared to the other known and available beverages or, carbonated drinks.

The whey based beverage can be produced at cheaper rate because of low capital investment. Also these have potential market. Hence the soft drink production plant can be an extension to cheese manufacturing dairy industries. Such a plant can be incorporated in dairy industry in future expansion advantageously as it resolves the problem of effluent disposal. It gives thirst quenching properties. Whey contains 20% of protein found in milk. Whey is proven to be an extraordinary nutritional material. It is a complete protein with the presence of all essential and non-essential amino acids. Whey protein also possesses very high biological value that is more than those of soy protein, egg white and casein. Whey protein has highest naturally occurring branched chain amino acid content.

Due to the capital investment such as plant can be installed with large as well as small scale dairy industry and thereby increasing the profitability of dairy industry as a whole. In the beginning the products experience high

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competition in well establish soft drink industries. The product can be made popular in the market through public awareness of nutritional states of whey and competitive brand.

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References


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Vinderola, C., Reinheimer, J., 2000. In bifdobacteria and lactic starter bacteria in fermented dairy products. International Dairy Journal 10, 271–275.

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Manufacturing of Funtional Whey Beverage

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