Manufacturing of Funtional Whey Beverage

INTRODUCTION
1. 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
U.D.C.T.Amravati Page 2
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” .
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
'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.
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.
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.
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.
Table: 3.1. Approximate composition of Whey
Constituent Cheese Whey % Casein Whey %

Total Solid 6.4 6.5
Water 93.6 93.5
Fat 0.05 0.04
True Protein 0.55 0.55
NPN(non-protein nitrogen 0.18 0.18
Lactose 4.8 4.9
Ash (minerals) 0.5 0.8
Calcium 0.043 0.12
Phosphorus 0.040 0.065
Sodium 0.050 0.050
Potassium 0.16 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.
 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
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
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
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.
Ingredients of whey
beverage
4. IN GENERAL, INGREDIENTS OF WHEY BEVERAGES

4.1. Whey 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 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
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).
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. Availability

1 Annatto E160B Liquid/ Powder
2 Caratenaid E160A Liquid
3 Marri Gold E161B Paste
4 Beetroot/ Carrot E162 Liquid/ Powder
5 Saff - Liquid/ Powder
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 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
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)
Beverage
Formulation
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 85.43
Fructose 9.00
WPI 5.00
Phosphoric acid 0.37
Natural mango flavor 0.05
Yellow color 0.04
Potassium sorbate 0.04
Salt 0.04
Calcium Chloride 0.02
Potassium Chloride 0.01
Total 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).
Nutritional Content per 100 grams
Calories 50 kcal
Total Fat 0g
Saturated Fat 0g
Trans Fat 0g
Cholesterol 0 mg
Sodium 20 mg
Total Carbohydrate 9g
Dietary Fiber 0g
Sugars 9g
Protein 5g
Vitamin C 0g
Vitamin B1 0g
Vitamin B2 0g
Calcium 2g
2. Low pH Juice Drink with WPI (Whey Protein Isolated)
Ingredients Usage Level %

Water 80.73
High Fructose corn Syrup 9.40
WIP 4.70
Apple Juice Concentrate -70 Brix 4.70
Phosphoric Acid Solution-85% 0.35
Natural Berry Flour 0.10
Red Color 0.02
Total 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.
4. Heat to 90ºC (195°F) for 45 seconds.
Calories 60 kcal
Total Fat 0g
Saturated Fat 0g
Trans Fat 0g
Cholesterol 0 mg
Sodium 0 mg
Total Carbohydrate 11 g
Dietary Fiber 0g
Sugars 7g
Protein 4g
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 93.00
Granulated Sugar 4.70
WPC 80 1.40
Vanilla Extract 0.50
Mono & Diglycerides 0.20
Carrageenan 0.10
Tetrasodium pyrophosphate 0.10
Total 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).
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 kcal
Total Fat 1g
Saturated Fat 0.5 g
Trans Fat 0g
Cholesterol 5g
Sodium 110 mg
Dietary Fiber 0g
Sugars 9g
Protein 4g
Vitamin C 0g
Vitamin B1 0g
Vitamin B2 0g
Calcium 120 g
4. Low Sugar Drink with WPC 80
Ingredients Usage Level (%)

Water 90.62
WPC 80 5.29
Cream 2.11
Pectin 1.37
Mango Flour 0.20
Sucralose 0.10
Red Colour 0.02
Yellow Colour 0.01
Total 100
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 sweetener, flavor and
colors.
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).
Nutritional Content per 100 grams:
Calories 30g
Total Fat 1g
Saturated Fat 0.5g
Trans Fat 0g
Cholesterol 5g
Sodium 15g
Dietary Fiber 0g
Sugars 0g
Protein 4g
Vitamin C 3.6g
Vitamin B1 0.02g
Vitamin B2 0.05g
Calcium 24g
5. Juice Drink with WPC 80(Whey Protein Concentrate)
Ingredient Usage Level (%)

Water 79.57
Granulated Sugar 8.33
WPC 80 5.20
Juice concentrate ,Pectin 5.09
Pectin 1.30
Milk Calcium 0.31
Total 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
Total Fat 0g
Saturated Fat 0g
Trans Fat 0g
Cholesterol 0g
Sodium 20g
Dietary Fiber 0g
Sugars 11g
Protein 4g
Vitamin C 9g
Vitamin B1 0.02g
Vitamin B2 0.05g
Calcium 96g
(Steve Rittmanic, 2006)
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)
hR.
92e8spPuvy&bfW
10-5ColdStraingficFtizHm
6.3. Whey Processing:
Fat Content 25-30%
By Ultrafiltration
Fig.6.1. Flow chart of whey processing (Ref. Mirjan Djuric 2004)
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.
Whey Beverage Manufacturing (In general):
Fig.6.2. Flow Chart of Beverage Manufacturing

(Ref. Article on Acido whey, 2010)
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.
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.
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.
WHEY PROTEIN CONCENTRATE RECEIVING OF MANGO

(35%)
SHORTENING OF FRUIT
RECONSTITUTE WPC WITH
DISTILLED WATER BY 10%
REMOVING OF CROWN
TRANSFERRED INTO
CUTTING INTO SMALL
INCUBETED AT 370C FOR

24hr PULPING IN MIXER
FERMENTATION PASSING INTO HAND
BOTTLING OF WHEY PASSING INTO HAND
BLENDING OF WHEY AND

MONGO JUICE (70:30)
DISSOLVED 2% CALCIUM LACTED
FORTIFICATION WITH SUGAR(PH 4.2-
FILTRATION AND CLARIFICATION
BOTTLING AND CARKING
ASEPTIC PACKAGING & STORAGE(8-100C)

Fig.7.1. Flow chart of Functional whey beverage formulation
(Ref. Micaela Pescuma, 2010)
Quality Control and

Testing
8. 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
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
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.
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.
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
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 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
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.
Machineries
9. Machineries
1. Fermenter
2. Centrifuge for separation of fat
3. Pasteurizer
4. Filling Machine
5. 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.
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.
1. Pasteurizer
Pasteurizer equipment is used for pasteurization of whey and fruit juice.

Pasteurization is done at 80-850C.
2. Filling Machine
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.
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.
Loss Occurs (Standardization,

500L of whey 450L whey
Pasteurization, sterilization) remains
0
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, 135L Fruit Juice

Filtration) remains
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
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)
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
Therefore,
(77519+23255.7+13645+1046.12) L = (100375+5090.82) L
105465.82L = (100375+5090.82) L
= 1, 00,000L (approx.)
Cost estimation
11. COST ESTIMATION
1. Fixed Capital:
A) Equipment Cost
Equipment
SN. Equipment Cost(Lakh)
1 Fermenter 50L
2 Pasteurizer 15L
3 Sterilizer 10L
4 Mixing Tank 0.8L
5 Storage Tank 8L
6 Inoculums Tank 12L
7 Laboratory Equipment 3L
8 Generator 10L
9 Labeling Machine 0.8L
10 Centrifuge 10L
Total 1,16,70,000Rs.
A) Direct Cost
F1 = Purchase equipment cost = 0.15

F2 = Instrumentation and control = 0.10
F3 = Piping (installed) = 0.15
F4 = Electrical (installed) = 0.15
F5 = Building (Including services) = 0.20
F6 = Utilities = 0.50
F7 = Yield Improvement = 0.05
F8 = Auxillary Building = 0.15
TOTAL = 1.45
Total Direct Cost = Total X Equipment Cost
=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
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.
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
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.
Plant layout
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.
Conclusion
14. 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
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.
References
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.
 Djurić, M., Carić, M., Milanović, S., Tekić, M., Panić, 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 using lactic acid bacteria, Int.
journal of food microbiology.2010Jun30, 141(1-2):73-81.Epub
2010Apr18.Journal of Dairy Science 74, 20–28.
 Horton, B.S, 1995.Whey 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 Microbiology 141 (2010)

73–81.
 Mljekarstvo, Whey based beverages-new generation of dairy products,

2008.
 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., J.F. Kennedy, D.N.Gandhi and k. Bunko, 2007. Bio-

utilization 0f whey for lactic acid production. Food 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 Journal of Food Chemistry ,Whey

utilization,57-58
 Singh S. Ladkani, B.G. Kumar, Mathur B.N.1994,Development of whey

based beverage Indian J. Dairy Science,147(7:585)
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Dairy J.18:695-704.
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browing inhibitors in mango butter –milk and whey beverages. Egy.J.
Dairy Sci. 23:11-29.
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Rittmanic, Nutritional Food and Beverage Development, Arizona, USA.
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Bifidobacterium bifidumand Lactobacillus acidophilus in the presence of
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Manufacturing of Funtional Whey Beverage

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Manufacturing of Funtional Whey Beverage

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INTRODUCTION

In order to provide assistance to Alma Creamery, the Kansas Department

1.1. Importance of whey Beverage

Beverages based on whey continue to receive a considerable amount of

Out of 85 million tones of global production 40% is still disposed as raw

The present investigation has been proposed to develop whey based

1.2. Whey Proteins

From a functional perspective, dried whey protein is appropriate for

1.3. Commercial Whey Protein Products

Given the advantages of whey protein, it has become a popular source of

A functional beverage can be defined as a drink product that is non-

Functional beverages have become popular due to its appeal to consumers

1.4. Health and wellness

Functional beverage companies are more aware of the ‘health conscious’

1.5. Weight management

With increased worries about obesity and its implications on health,

Aims and objective

2. Aims and objectives:

Whey constituted about 80-92% of the volume of milk used for

A functional liquid whey based beverage system was developed, keeping in

1. To develop the functional whey beverages based on whey which is

Because of it high organic matter content, milk whey constitutes a serious

Whey is the by-product of cheese and casein manufacture, being what

Table: 3.1. Approximate composition of Whey

Constituent Cheese Whey % Casein Whey %

Available literature indicates that whey beverage have been studied

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

 Research at University of Novi Sad, Faculty of Technology, studied that

 Research (Mljekarstvo, 2008) investigates the Whey based beverages-

 Another research had done in University of Novi Sad, Faculty of

 Comparative Analysis of Indian Paneer and Cheese Whey for

 Biology Department, Federal University of Lavras, 37200-000 Lavras,

4. IN GENERAL, INGREDIENTS OF WHEY BEVERAGES

The most important component of an acidified whey protein RTD (Ready

4.2. Non-protein ingredient considerations

Following are some of the other categories of ingredients frequently required or

Whey protein’s strong buffering capacity requires the use of considerable

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

2) Hydrochloric – a strong acid with less desirable palatability, but may be

4) Malic – a weaker acid similar to citric acid but useful as an adjunct to

2. Carbon Dioxide (carbonation)

Carbonation is featured in this section because it should indeed be considered an

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

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

Colors may be either artificial or natural, with light stability an important

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

Sr. No Name of colour EEC No. Availability

The stability and clarity of acidified whey protein beverages is believed to

As with any food or beverage product, vitamin must be chosen and

9. Stabilizers and Emulsifiers

Stabilizers and emulsifiers can be very important to neutral, shake-type

Acidified whey protein beverage formulas can include chemical

Whey protein drinks are considered high-value nutritional beverages, and

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

Ingredients Usage Level (%)

1. Reconstitute WPI in formula water (at ambient temperature) with a high

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).

Nutritional Content per 100 grams

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