Manufacturing of Funtional Whey Beverage

INTRODUCTION

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

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 discharge of whey into local waterways, waterways, the ocean, or as a component component in ani mal 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 processin g 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. (Smith (Smithers, ers, 2008) 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, Manhattan, KS), the proposed beverage would contin ue to be formulated formulated with a liquid whey base, but with the added value of carbonation.

U.D.C.T.Amravati U.D.C.T.Amravati

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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 1.1.

Importance of whey Beverage

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 improper nutrition leading to deficienc ies 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 consumption . (Horton, B.S.1995) The present investigation has been proposed to develop whey based functional functional whey beverage by mixing appropria appropria te 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 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, ³complete source´ of protein. More specifically, whey proteins are a rich source of branched chain amino acids (BCAAs), containing the highest known levels levels of any natu ral food source. BCAAs are important 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 ³consuming supplemental whey protein weighed less and put on less body fat compared to individuals who consumed consumed a calorie-equivalent carbohydrate carbohydrate supplement´ supplement´ . U.D.C.T.Amravati U.D.C.T.Amravati

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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 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 imparted from the whey protein lends lends itself well t o 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 whey protein beverage beverage would either either be far above or b elow 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.

C ommercial ommercial 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, ultrafiltra tion, and/or hydrolysis hydrolysi s treatments. treatments. The intent of this research was to utilize the unprocess unprocess ed 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 functional beverage can be defined as a drink product that is non alcoholic, alcoholic, ready to drink and includes in its formulation formulation non -traditional -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, others, are some of the product segments rolled out as function function al beverages in the market space.


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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage Functional beverages have become popular due to its appeal to consumers who are seeking specific health benefits in their foods and beverages with their 'healthiness-on-the-go' 'healthiness-on-the-go' idea. Both convenience and health have 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 improved immunity and digestion, digestion, joint health, satiety, and energy energy boosting. (Sharma & vahati K.L., 1999) 1.4.

Health

and wellness

Functional beverage companies are more aware of the µhealth conscious¶ individuals and have introduced functional beverages with less sugar and therefore less calories. 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 B 3, 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 calorie content (although (although taste is a nother matter) 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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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage

A IMS IMS A ND

OBJECTIVE


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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 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 conversion into channa, paneer, cheese cheese and casein. It retained about 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 containing valuab le nutrients are dumped 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 product was evaluated based on on physical, microbiological microbiological,, an d sensory characteristics. With minor formulation changes, this beverage could realistically and and competitively competitively exist in today¶s marketplace. 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 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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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage Because of it high organic organic matter content, milk whey constitutes constitutes a ser ious 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 fermented dairy products. 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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L ITER ITER A A TURE

SURVEY


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LITERATURE SURVEY

Growing concern over pollution and environment control has renewed the pressure on the cheese cheese manufacture manufacture to stop dumping whey into streams and and municipal sewage system. Consequently, the search has begun again for new methods to use whey. whey. In light of growing global food 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 beverage in human nutrition, especially for therapeutic 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 th many doctors for various diseases, by the middle 19 century, whey cures reached a high point point with the establishment of over 400 whey houses 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 whey per day . Whey is the by-product of cheese and casein manufacture, manufacture, b eing 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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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage T able: able: 3.1. Approximate composition of Whey C onstituent C heese C asein onstituent heese Whey % asein Whey % Total Solid 6.4 6.5 Water 93.6 93.5 Fat 0.05 True Protein 0.55 0.55 NPN(non-protein NPN(non-p rotein nitrogen 0.18 0.18 Lactose 4.8 Ash (minerals) 0.5 0.8 Calcium 0.043 0.12 Phosphorus Phosphorus 0.040 Sodium 0.050 0.050 Potassium 0.16 Chloride 0.11 0.11 Lactic 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. y

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 characterist ic 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


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0.04

4.9

0.065 0.16

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage depends on the sucrose content, while the quality of blends with peach 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.

y

Research (Mljekarstvo, 2008) investigates the Whey based beveragesnew 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 products obtained by mixing mixing native sweet, diluted or aci d 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 produce functional functional beverage with high nutritional nutritional value an d 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 produ ction of alcoholic beverages beverages due to the fact t hat 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.


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

y

Another research had done in University of Novi Sad, Faculty of Technology, and Navi Sad, Serbia. They investigated the manufacture of milk-based beverages obtained by Kombucha application. Local Kombucha Kombucha culture was grown up on three three substrates: sweetened sweetened bla ck 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 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 electrolyte drink. The slight differences differences attributed, among various various parameters can be mainly due the difference in manufacturing process. Our results indicated significantly higher concentration of sodium, potassium, potassium, calcium and chlo ride contents in paneer whey whey than cheese whey. The analysis is important as paneer whey can be utilized more efficiently otherwise creating environmental pollution especially in India.


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

Biology Department, 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 morphological 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, 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 robustness is determinant determinant for future implementation implementation of whey based kefir beverages.


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INGREDIENTS OF WHEY BEVER A A GE GE


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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 4. 4.1.

IN GENERAL, INGREDIENTS OF Whey

WHEY

BEVERAGES

protein ingredient considerations

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 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 concentration to the presence p resence 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 stages o f the development development process. Many U.S. suppliers offer guidance, guidance, typical formulations 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 beverage contains a protein level on the high end of the practical range. range. Again, close and early collaboration with a U.S. supplier is an important success factor.


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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 4.2.

Non-protein ingredient considerations

Following are some of the other categories of ingred in gredients ients 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.

± 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. 2) Hydrochloric

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

3)

extreme tartness imparted when used at high levels. acid similar to citric acid but useful as an adjunct adjunct to Malic ± a weaker acid formulas with apple or berry flavors, due to its natural presence in those fruits.

4)

2. C arbon arbon 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 interest in improving 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


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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage beverages beverages are also less prone to the flavor adsorptive adsorptive effects of other proteins in beverages, a phenomenon which would require heavier flavor usage (and cost). 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 selection of sweeteners can impact mouth feel and protein stability in a formula specific manner. However, the choice of a sweetener sweetener is usually u sually 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 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 a re used in whey whey beverage formulatio n:

Sr. No Name of colour 1 Annatto 2 Caratenaid Caratenaid 3 Marri Gold 4 Beetroot/ Carrot 5 Saff


C EE No.

E160B E160A E161B E162 -

Availability Liquid/ Powder Liquid Paste Liquid/ Powder Liquid/ Powder

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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 7 . Fruit Juices

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 amount of mineral mineral ions, such as sodium or calcium, 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 aggregation in thermally processed whey beverages, thus decreasing stability. 8.

V itamins itamins

As with any food or beverage product, product, vitamin vitamin must be chosen and formulated formulated according to their compatibility compatibility with the overall system .Most watersoluble 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 translucent bottle. Ingredient interactions interactions should also be considered. Generally vitamin B1, Vitamin B2, Vitamin C etc. 9. Stabilizers and Emulsifiers

Stabilizers Stabilizers and emulsifiers can be very important important to neutral, shake -type beverages, especially when mixtures of proteins and/or cocoa powder are used. Carrageenan, 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 between 3.5 and 4.6 to protect 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 pyrophosphate are commonly commonly used in neutral pH beverages 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. U.D.C.T.Amravati U.D.C.T.Amravati

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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 9. Preservatives

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. spoilage. 10. Nutraceuticals 10. Nutraceuticals

Whey protein drinks are considered high-value nutritional beverages, and are often fortified with additional nutritional nutritional components components such as plant sterols to lower cholesterol, lutein to improve improve eye health, orma-huang orma-huang or guarana which are reported to boost energy. Live and active cultures 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, Rittmanic, 2006)


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BEVER A A GE GE FORMULA TION


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

Water Fructose WPI Phosphoric Phosphoric acid Natural mango mango flavor Yellow color Potassium sorbate Salt Calcium Chloride Chloride Potassium Chloride Total

Usage Level (%) 85.43 9.00 5.00 0.37

0.05 0.04 0.04 0.04 0.02 0.01 100

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 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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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage Nutritional Content per 100 grams

50 kcal 0g 0g 0g 0 mg 20 mg 9g 0g 9g 5g 0g 0g 0g 2g

Calories Total Fat Saturated Saturated Fat Trans Fat Cholesterol Sodium Total Carbohydrate Dietary Fiber Sugars Protein Vitamin C Vitamin B1 Vitamin B2 Calcium

2.

Low pH Juice Drink with

WPI (Whey

Ingredients Water High Fructose corn Syrup WIP Apple Juice Concentrate -70 Brix Phosphoric Phosphoric Acid Solution-85% Natural Berry Flour Red Color Total

Protein Isolated)

Usage Level % 80.73 9.40 4.70 4.70 0.35 0.10 0.02 100

Procedure:

1. Reconstitute WPI in formula water (at ambient temperature)with 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 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). U.D.C.T.Amravati U.D.C.T.Amravati

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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage Nutritional Content per 100 grams Calories Total Fat Saturated Fat Trans Fat Cholesterol Sodium Total Carbohydrate Dietary Fiber Sugars Protein Vitamin C Vitamin B1 Vitamin B2 Calcium

3.

Meal Replacement Drink with

Ingredients Skim milk Granulated Granulated Sugar WPC 80 Vanilla Extract Mono & Diglycerides Carrageenan Tetrasodium pyrophosphate Total

60 kcal 0g 0g 0g 0 mg 0 mg 11 g 0g 7g 4g 0 mg 0 mg 0 mg 0 mg

WPC 80( Whey

Protein Concentrate)

Usage Level(%) 93.00 4.70 1.40 0.50 0.20 0.10 0.10 100

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.


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Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 5. Heat to 85ºC (185°F). 6. Homogenize: first stage at 250 Bar (24.82MPa, 250kg/cm2 or 3600psi) and second stage stage at 48 Bar (4.82MPa, (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. Nutritional Content per 100 grams

60 kcal 1g 0.5 g 0g 5g 110 mg 9g 0g 9g 4g 0g 0g 0g 120 g

Calories Total Fat Saturated Saturated Fat Trans Fat Cholesterol Sodium Total Carbohydrate Dietary Fiber Sugars Protein Vitamin C Vitamin B1 Vitamin B2 Calcium

4.

Low Sugar Drink with

WPC 80

Ingredients

Water WPC 80 Cream Pectin Phosphoric Phospho ric acid Mango Flour Sucralose Red Colour Yellow Colour Total


Usage Level (%)

90.62 5.29 2.11 1.37 0.28 0.20 0.10 0.02 0.01 100

Page 25


Procedure:

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 sweeten er, 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 Total Fat Saturated Saturated Fat Trans Fat Cholesterol Sodium Total Carbohydrate Dietary Fiber Sugars Protein Vitamin C Vitamin B1 Vitamin B2 Calcium


30g 1g 0.5g 0g 5g 15g 2g 0g 0g 4g 3.6g 0.02g 0.05g 24g

Page 26

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 5. Juice Drink with WPC 80( Whey Protein Concentrate) Ingredient Water Granulated Granulated Sugar WPC 80 Juice concentrate concentrate ,Pectin Pectin Milk Calcium Phosphoric Phosphoric acid Total

Usage Level (%) 79.57 8.33 5.20 5.09 1.30 0.31 0.20 100

Procedure:

1. Hydrate stabilizer and sugar in half of the formula water at 85ºC (185°F) and let swell s well for10minutes. 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 concentrate and add colors for desired tint. 9. Fill containers and cool to 4ºC (40°F).


Page 27

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage Nutritional Content per 100 grams Calories Total Fat Saturated Fat Trans Fat Cholesterol Sodium Total Carbohydrate Dietary Fiber Sugars Protein Vitamin C Vitamin B1 Vitamin B2 Calcium (Steve Rittmanic, 2006)


70g 0g 0g 0g 0g 20g 12g 0g 11g 4g 9g 0.02g 0.05g 96g

Page 28


MA TERIAL A ND


METHODS

Page 29

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 6. 6.1.

MATERIALS AND METHODS

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)


Page 30

Manu Manu acturi acturing ng Of 1, 00,000 Litr / ay Func Functi ti nal Wh Wh y Bever Beverage 6.3. Whey Processi Processi Recieving of Whey

Stored temporarily at about 500C

Casein fines recovary & fat separation

Fat Content 25-30%

Pasteurisation at 850C For 10min.

Sterilization 90-920C For 5min

Homogenization

Filtration & Clarification

By Ultrafiltrat U ltrafiltration ion

Cooling at 10-150 C

Cold Storage

Fi .6.1. Fl Flow chart of whey processi processi U.D.C.T. Amr Amr 



¡

¢

i

(Ref. Mi Mirjan Djuri Djur ic 2004) Pa g 31 £

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 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 incubation tank pre-adjusted to the specified temperature. 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 whey is inoculate i noculated d with a pure and active culture at a desired level. level. After inoculation when the acidity of the whey reaches reaches at the

desired level the fermented whey is cooled and filtered through the filter press. 4.

Fortification with sugar and flavor:

Filtered fermented 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 prepared whey beverage 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 beverage should be pasteurized before packaging or alternatively pasteurized in the container.


Page 32

Manu Manufacturi facturing ng Of 1, 00,000 Litre Litre/ ay Func Functi ti nal Wh Whey Bever Beverage Whey Beverage Manufacturi Manufactur ing (In general general):

Collection of whey

tand ta ndar ardi di at atio ion n terilii at teril atio ion n and and Pa teuri ation at 850C for 10min Cooling at (15200C) Blending With Fruit Juice Fortification With ugar and E ence Filtration And Clarification Flav Fl avori oring( ng( Adj Adju u t PH 3.6-4.2 ) Bottling and Corcking Packaging

Cold torage

Fig.6.2. Flow Chart of B of Beverage Manufacturi Manufactur ing (R U.D.C.T. Amr Amr avat avat i i

Ar ti ti l on Aci Aci o whey, 2010) Pa g 33 ¤


FORMULA TION OF FUNCTIONAL FERMENTED WHEY A GE BA SED BEVER A GE

USING LA CTIC CTIC A CID CID BA CTERI CTERIA


Page 34


FORMULATION OF FUNCTIONAL FERMENTED WHEYBASED BEVERAGE USING LACTIC ACID BACTERIA(Case Study)

7.1.

Preparation of media:

1. Microorganisms and media

The strains strains L. acidophilus CRL 636, L. 636, L. delbrueckii subsp. subsp. Bulgaricus Bulgaricus CRL 656 and Streptoco Streptococcus ccus 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 í20 °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 deterioratin deteriorating g 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), aureus), violet red bile lactose agar (VRBA, for total coliforms), plate count agar (PCA, for mesophilic microorganisms), microorg anisms), and potato dextrose 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.


Page 35


Fermentation conditions

Cultures were transferred twice in WPC35 prior to experimental use; 16 h old cultures (2% v/v) were used as inoculums individually, 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. incubati on. Cell viability was determined by plating appropriate appropriate dilutions of the cultures in MRS agar (MRS Britania, Buenos Aires, Argentina, plus 15 g/l agar). To determine determine the viable viable cell count of the L. acidoph acidophilus ilus strain in the mixed culture, culture, 1.5% (w/v) bile salt (Sigma Chemical CO, St. Louis, USA) was added to MRS agar (Vinderola and Reinheimer, 2000). 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 of L. delbrueckii subsp. bulgaricus and S. thermophilus in yogurt. (Vinderola ( Vinderola and Reinheimer 1999). 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 every 2 h during the first 12 h and and after 24 h incubation.


Page 36


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 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 ( 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 activity protein degradation degradation we re determined determ ined after 0, 7, 14, 21 and 28 days of storage .


Page 37

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage RECEIVING OF MANGO FRUIT

WHEY PROTEIN CONCENTRATE (35%)

SHORTENING OF FRUIT RECONSTITUTE WPC WITH DISTILLED WATER BY REMOVING OF CROWN TRANSFERRED INTO CULTURE CUTTING INTO SMALL PIECES INCUBETED AT 37 0 C FOR 24hr

PULPING IN MIXER

FERMENTATION

PASSING INTO HAND PULPER

BOTTLING OF WHEY

PASSING INTO HAND PULPER

BLENDING OF WHEY AND MONGO JUICE (70:30)

DISSOLVED 2% CALCIUM LACTED

H

FORTIFICATION WITH SUGAR(P 4.2-5)

FILTRATION AND CLARIFICATION

BOTTLING AND CARKING

0

ASEPTIC PACKAGING & STORAGE(8-10 C)

Fig.7.1. Flow chart of Functional whey beverage formulation

(Ref. Micaela Pescuma Pescuma , 2010) 2010) U.D.C.T.Amravati U.D.C.T.Amravati

Page 38


QUAL ITY ITY CONTROL A ND


TESTING

Page 39

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 8. QUALITY CONTROL AND TESTING

1. Analysis of metabolites

Sugar Sugar content (lactose, galactose and glucose) glucose) and organic acids (lactic, acetic, and formic) formic) production production were analyzed during during fermentation 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 proteolytic activity of LAB was measured 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 -phthaldialdehyde (OPA) ( 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 (previously dissolved in 1 ml methanol), methanol), 100 l of 2mercaptoethanol (Merck, Darmstadt, Germany) and distilled water up to a 50 ml final volume. Fermented Fermented samples were incubated with 0.75 mol/l trichloroacetic trichloroacetic acid (Sigma Chemical Chemical Co) at a sample: trichloroacetic 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 g leucine (Leu) released/ml using a standard curve of L-leucine (BDH Chemicals Ltd Poole, England).


Page 40


Free amino acid determination

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 l of WPC35 with 2% (w/v) of SDS (dissolved in 0.4 mol/l sodium borate buffer, pH 9.5) and 200 l of the OPA methalonic solution. The mixture was shaken, incubated for 1 min and the reaction was stopped by adding 400 l of 0.1 mol/l sodium phosphate buffer (pH 4.0) and filtered through 0.2 m nylon 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 NE,USA 5 m, 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. 20min. Absorbance was recorded at at 305±395 and 430±460 nm excitation and emission wavelengths, respectively. The injection volume of derivative amino acids was 10 l. The HPLC was coupled with the software Chem. Research 150 Data System 3.0.2. All the amino acids, except proline, cystein and methionine, methionine, were determined under the assayed conditions. conditions. Amino acid concentration was expressed in g/ml. 4. Hydrolysis

of  -lactoglobulin in

WPC35

Degradation of -lactoglobulin was monitored by RP-HPLC using a Knauer Smartline System Syste m (Manager 5000, pump pump 1000) with a UV detector (2000) fitted with a C18 column (Pursuit 4.6×250 4.6×250 mm, 300 A, 5 m, 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 U.D.C.T.Amravati U.D.C.T.Amravati

Page 41

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 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 HPLC were prepared as follows: WPC35 samples (fermented and nonfermented) were mixed 1:1 with reduction buffer containing urea and 20 mole/l dithiothreitol dithiot hreitol (DTT) and incubated for 60 min at 30 °C. Prior to injectio injection n 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 (non -fermen -fermented ted sample). 2.8. Strains compatibility Strains compatibility was evaluated by the plate diffusion assay (Parente and Zottola, Zottola, 1991). Briefly, Briefly, overnight cu ltures 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 l of the cell suspension on the agar. After overlay solidification, 5 mm diameter wells made with sterilized plastic straws were inoculated with 60 l 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 results were expressed as mean values with standard standard deviations. Statistical analyses analyses were perform performed ed using usin g 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 content was measured using an Ab be Mark2 Refractometer. 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


Page 42

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage measurements were taken from both the uncarbonated and carbonated samples and then averaged. 8 . C olor olor

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 µa*¶ value indicates redness, and a negative negative µa* ¶ value indicates greenness. A positive µb*¶ value represents represents yellowness and a negative negative µb*¶ value repr esents 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. Procedure:

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 U.D.C.T.Amravati U.D.C.T.Amravati

Page 43

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 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 evaporate off the solvent i n the flask on a water bath. Remove the traces of the 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 fat in a 250 ml conical conical flask and add 50 ml. ml. of mixed benzene -alcohol-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. C arbonation arbonation 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 accompanying gauge. gauge. The device¶s thermometer 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 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 beverage products. One milliliter of the product, 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 previously stated serial dilutio dilutio ns, was plated on 3M Petrifil Petrifil (ECC Plates, 3M, and MN) and then incubated at 32°C 1°C for 48h 3h. U.D.C.T.Amravati U.D.C.T.Amravati

Page 44

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 12. Sensory Sensory

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 anchored on the left by 1 (³dislike extremely´) extremely´) and on t he right by 9 (³like extremely´).

13. Nutrition 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.


Page 45


MA CHINERIES CHINERIES


Page 46

Manu Manufacturi facturing ng Of 1, 00,000 Litre Litre/ ay Func Functi ti nal Wh Whey Bever Beverage 9.

1. 2. 3. 4. 5.

Machi Machiner neriies Ferment Fermenter Cent entr i e for separati separation on of fat fat Past Pasteur i er Filling illing Machi achine Col Storage-R orage-R efr efr igerat gerator

1. Fermenter:

(R ef.At ef.Athor,Dr.J.N.de Wit Wit,, 2009) Ferment Fermenters that hat is procured from reli re liab ablle manufact manufacturers. The range of Ferment Fermenters offered by us is renowned for it for itss sturdy const constructi ruc tion on and durabilit durab ility. y. Fur ther, the range of Ferment Fermenters offered by us does not no t react reac t with ith chemi chemical ca ls, thereby offer ing high utility tility and effecti effec ti eness to the clien lientts. Our range of Ferment Fermenters is avail availab ablle in var ious capaciti capacities es to ensure hi high utilit utility y to the cli c lien entts. The hi high qualit qua lity y and effecti effec tive ve performance of these Ferment Fermenters have made them the preferred choi choice across indust ndustr ies gl globall obally. y. U.D.C.T. Amr Amr avat avat i i

Pa g 47 ¥

Manu Manufacturi facturing ng Of 1, 00,000 Litre Litre/ ay Func Functi ti nal Wh Whey Bever Beverage

2. Centri Centr ifuge:

(R ef.At ef.Athor,Dr.J.N.de Wit Wit,, 2009) The Cent entr ifuges that hat is widel dely used to separat separate subst substances of di different fferen t densiti densities. es. The Cent entr ifuges offered by us have been fabr icat ca ted usi using high grade raw mat mater ial. Fur ther, we foll follow ow an ext extensi ens ive qualit quality y management management program to ensure compli compliance ance of our range wit w ith h var ious internati erna tiona onall qualit quality y standards. The range of Cent entr ifuges offered by us is renowned for its its noi noisel seless and effecti effective ve performance, and is wi w idel dely used in chemi chemical ca l as well well as pharmaceuti pharmaceutica call indust ndustr ies. Si Since our range of Cent entr ifuges has been manufact manufactured as per internati ernationa onall safet safety norms it offers enhanced safet safe ty.

U.D.C.T. Amr Amr avat avat i i

Pa g 48 ¦

Manu Manufacturi facturing ng Of 1, 00,000 Litre Litre/ ay Func Functi ti nal Wh Whey Bever Beverage

1. Pasteuri Pasteur i er

(R ef.At ef.Athor,Dr.J.N.de hor,Dr.J.N.de Wit, it, 2009) Past Pasteur i er equi equi pment pment is used for past pasteur i ation tion of whey and fruit fru it j ju uice. 0 Past Pasteur i a tion tion is done at a t 80-85 C. 2. Filling illi ng Machi Machine

(R ef.At ef.Athor,Dr.J.N.de Wit Wit,, 2009)

U.D.C.T. Amr Amr avat avat i i

Pa g 49 §

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage The bottle filling machine is capable to fill 60 /120 machines 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.


Page 50


MA TERIAL BALA NCE


Page 51


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, Standardization, sterilization and pasteurization. pasteurization.

500L of whey

Loss Occurs

(Standardizati (Standardization,

Pasteurization, sterilization)

450L

whey remains

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. 150L of whey

Loss

Occurs (Clarification,

Filtration)

3.

135L Fruit Juice remains

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 U.D.C.T.Amravati U.D.C.T.Amravati

Page 52


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

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 1.

Whey

Beverage:

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)

Page 53


Addition of (1) & (2) 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*(5/10 105658*(5/100 0 ) = 5282.9L Therefore, 105658 ± 5282.9 = 100375.1L = 1, 00,000L (nearly)


Page 54

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage Material balance for the manufacturing of whey beverage is as follows: Feed

=

Product + Waste

Therefore, (77519+23255.7+13645+1046.12) L

=

(100375+5090.82) L

105465.82L

=

(100375+5090.82) L

=

1, 00,000L (approx.)


Page 55


COST ESTIMA TION


Page 56

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 11. COST ESTIMATION 1. Fixed Capital: A) Equipment Cost Equipment SN.

1 2 3 4 5 6 7 8 9 10

Equipment

Cost(Lakh)

Fermenter Fermenter Pasteurizer Pasteurizer Sterilizer Mixing Tank Storage Storage Tank Inoculums Inoculums Tank Laboratory Laboratory Equipment Equipment Generator Generator Labeling Machine Machine Centrifuge Centrifuge Total

50L 15L 10L 0.8L 8L 12L 3L 10L 0.8L 10L 1,16,70,000Rs.

A) Direct Cost F1 F2 F3 F4 F5 F6 F7 F8

= = = = = = = =

Purchase equipment cost Instrumentation Instru mentation and control Piping (installed) Electrical (installed) Building (Including services) Utilities Yield Improvement Auxillary Auxillary Building TOTAL


= = = = = = = =

0.15 0.10 0.15 0.15 0.20 0.50 0.05 0.15

=

1.45

Page 57

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage Total Direct Cost = Total X Equipment Cost =1.45 X 1, 16, 70,000 =16, 92, 1500 Rs.

B) Indirect cost F9

=

F10

=

F11 F12

Engineering & Supervision

=

0.20

Construction expenses

=

0.15

=

Contractor Fees

=

0.10

=

Contingency Contingency

=

0.15

=

0.60

Total

Indirect Cost

Total Fixed Cost

Working Capital


=

0.60 X 1, 16, 70,000

=

70, 02,000

=

Equipment Cost + Direct Cost + Indirect Cost

=

3, 55, 93,500Rs.

=

15% of fixed cost

=

0.15 X 3, 55, 93,500

=

53, 39,025Rs.

Page 58

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage C) Land Cost

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 2. Sugar 3. Fruit juice 4. Colour and Essence

Total

= = = =

=

5, 42,633Rs. 27,290Rs. 6, 97,671Rs. (as per 30Rs./lit) 30Rs./li t) 32,406Rs.

13, 00,000Rs.

b) Miscellaneous Material

= =

10% of Working capital 5, 33,902.5Rs.

c) Utilities Utilitie s (Fresh Water, Electricity)

= =

10% of fixed cost 35, 59,350Rs.

d) Transportation Cost

= =

5% of raw material 65,000Rs.


Page 59

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

=

(a) + (b) + (c) + (d)

=

54, 58,252.5Rs.

= = = = = = = =

10% of fixed cost 35, 59,350.0Rs. 52, 00,000Rs. 60% 0f fixed cost 2, 13, 56,100Rs. 1% of foxed cost 3, 55,935Rs. 2% of fixed cost 7, 11,870Rs.

=

2, 84, 83,255Rs.

F) Other Fixed Cost

a) Maintenance b) Operating labour and staff c) Plant over head d) Insurance e) Royalty

Total

Direct production cost


=

Variable cost + Other Fixed cost

=

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

=

3, 39, 41,507.5Rs.

Page 60


G) Sale Income

Whey Beverage produced per year Price of Whey Beverage

= 1, 00,000 L/ yr. = (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

b) Depreciation Cost

= =

5, 45, 00,00000,000 - 3, 39, 41,507.5

=

2, 05, 58,492.5 Rs.

= =

c) Taxable Income

d) Tax on Profit


10% of investment 20, 55,849.5Rs.

=

Gross return ± depreciation cost

=

2, 05, 58,492.5 ± 20, 55, 849

=

1, 85, 02,643.25Rs.

= =

e) Net Profit

sale income ± production cost

50% of taxable income 92, 51,321.625Rs.

= Gross Gross return± Depreciation cost-Tax cost -Tax = 2, 05, 58,492.5-2, 58,492.5 -2, 05, 58,449.25-92, 58,449.25 -92, 51,321.1625 = 92, 51,321Rs.

Page 61


PLA NT LA YOUT


Page 62

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

This layout can play an important part in determining const ruction 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 quality of products to be produced . 3. Type of process and product control. 4. Operational Operational convenience and acces sibility. 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 10.Roads and railroads.


Page 63


CONCL USION USION


Page 64

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 14. Conclusion Whey is a liquid product separated out from milk in the preparation of paneer and cheese. 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 beverage unlike the other carbonated 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 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 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 complete protein with the presence 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.


Page 65

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage 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 competition in well establish soft drink industries. The product can be made popular popular in the market through public awareness awareness of nutr itional states of whey and competitive brand.


Page 66


REFERENCES


Page 67

Manufacturing Of 1, 00,000 Litre/day Functional Whey Beverage References:  Anon. 1968.Dairy drink from cheese whey.Amer.Dairy Rev.30 (8):52 

CODEX STAN 192-1995. (Rev 2-1999). Sección 5.2. Norma general Del Codex para los aditivos alimentarios-preámbulo.



Djuri, M., Cari, M., Milanovi, S., Teki, M., Pani, M., 2004. Development of whey based beverages. European Food Research and Technology 219, 321±328.



Dr.Jde Wit.N., Consultant Food Protein, Renkum, Netherland, Lecturer Handbook on whey and whey products drink by Nupur Goyal and D.N.Gandhi, Journal of Dairy & Food Science, 70-72,2009.



Encyclopedia of food science, Food Technology & Nutrition, page no.4900-4491



Functional fermented whey based beverage Functional beverage usi ng lactic acid bacteria, Int. journal journal of o f food microbiology.20 microb iology.2010Jun3 10Jun30, 0, 141(1 -2):73-81.Ep -2):73-81.Epub ub 2010Apr18.Journal of Dairy Science 74, 20±28.



Horton,, B.S, Horton B. S, 1995.Whey processing processing and utilization.Bill.IDF, 308:2 -6.



Karina Teixeira Magalhaes, Biology Department, Federal University of Lavras, 37200-000 Lavras, MG, and Brazil have done the Production of fermented cheese whey-based beverage using kefir grains as starter culture: Evaluation of morphological and microbial variations. 2010.



Karina Teixeirira Magalhaes 2010, Journal of Bioresource Technology, Biology Department, Federal University of Lavras.



Marjana Djuric, Development of whey based beverages, panic Faculty of Technology, University of Navi sad, 2004.



Micaela Pescuma International Journal of Food Food Microbi ology 141 (2010) 73±81.



Mljekarstvo, Whey based beverages-new generation of dairy products, 2008.


Page 68




National research development of Acido whey, Research paper of government of India enterprise, Zamroodpur Community Centre, New Delhi, 2010.



Nupur Goyal and D.N.Gandhi, Comparative Analysis of Indian Paneer and Cheese Whey for Electrolyte Whey Drink, Dairy Microbiology Division, National Dairy Research Institute, Karnal, India.



Panesar, P.S., Panesar, P. S., J.F. Kennedy, D.N.Gandhi D.N.Gandhi and k. Bunko, 2007. Bio utilization utilizatio n 0f 0 f whey for lactic acid production. production. Food chem., chem., 105:1 -14



Parente, E., Zottola, E.A., 1991. Growth of thermophilic starters in whey permeates media. Raymond Kassatly, Kansas State University, Manhattan, KS.



Ranganna, S.1968. Handbook of Analysis and Quality Control for Fruit and Vegetable Products.



Rodomir V. Malbasa, 2009 utilization,57-58

Journal of Food Chemistry ,Whey



Singh S. Ladkani, B.G. Kumar, Mathur B.N.1994,Development of whey based beverage Indian J. Dairy Science,147(7:585)



Smither, G.W.2008. Whey and whey protein from gutter-to-gold. Int. Dairy J.18:695-704.



Soliman, M.A., E.A. Emave and A.E. Okasha, 1995.Studies on the browing inhibitors in mango butter ±milk and whey beverages. Egy.J. Dairy Sci. 23:11-29.



Steve Rittmanice,2006, U.S Whey protein in RTD Beverage by Steve Rittmanic, Nutritional Food and Beverage Development, Arizona, USA.



Vinderola, C., Reinheimer, J., 1999. Culture media for the enumeration of Bifidobacterium Bifidobacteriu m bifidumand Lactobacillus acidophilus in the presence of yoghurt yoghu rt bacteria. International Dairy Journal 9, 497 ±505.


Page 69

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

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


Page 70

Manufacturing of Funtional Whey Beverage

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