2.

0 REVIEW OF LITERATURE
Whey is a nutritious and valuable by-product obtained during manufacture of cheese,
chhana, paneer, casein, etc. Many attempts have been done on utilization of whey in the
formulation of various dairy products but, still there is a lot of scope to explore the
possibilities for its utilization. The disposal of whey creates a problem to environment. In
today’s generation many people are suffering with lactose intolerance. Therefore, the
utilization of Lactose Hydrolyzed Paneer Whey as a diluent is used in the preparation of
popular fermented drink like Lassi beverages is one of the best options for lactose
intolerance people.
The information with respect to significance of whey and utilization for the
production of fermented beverages has been critically reviewed in this chapter.

2.1 PANEER
Paneer is a South Asian variety of soft cheese obtained by acid and heat
coagulation of milk. It is a non-fermentative, non-renneted, non-melting and unripened
type of cheese. Paneer is popular throughout South Asia, used in raw form or in
preparation of several varieties of culinary dishes and snacks. The production of paneer is
now spreading throughout the world. The ability of paneer to be deep fried is one feature
that has led to its wider acceptance and a favourite for making snacks, pakoras or fried
paneer chunks (Aneja, 2007). Good quality paneer is characterized by a marble white
colour, sweetish, mildly acidic taste, nutty flavour, spongy body and closely knit, smooth
texture.

According FSSAI, (2006) Act, Paneer means the product obtained from the cow or
buffalo milk or a combination thereof by precipitation with sour milk, lactic acid or citric
acid. It shall not contain more than 70.0 per cent moisture and the milk fat content shall not
be less than 50.0 per cent of the dry matter. Milk solids may also be used in preparation of
this product.
 Review of Literature

India has witnessed a remarkable growth in milk production during the last few decades
due to the success of the Operation Flood Programme, which is one of the world’s largest
and successful integrated dairy development programs initiated in 1970s. The world milk
production in 2012 is forecast to grow by 3.0 per cent to 750 million tonnes. India is the
world’s largest milk producing country, with an output of 132 million tonnes (FAO, 2012).

2.2 WHEY
Whey is the major byproduct of the dairy industry obtained during the preparation
of different varieties of cheese, chhana, paneer and casein. Whey has been viewed recently
as one of the major disposal problems of the dairy industry. Whey is a serious pollutant as
it imposes a very high BOD of 30,000- 50,000 mg/L and chemical oxygen demand of
60,000-80,000 mg/L. Discarding of whey also constitutes a significant loss of potential
nutrients and energy and has been looked upon seriously by the environmentalists and
technologists due to its potent polluting strength. In addition the dairy industry suffers
from an economic blow due to several treatment costs that are required. Now-a-days it is
no longer considered a waste product but the treasure house of nutritionally rich whey
proteins and the modern technology enabling them to recover economically in their native
form. Thus utilization in the human food chain is now being predominantly favored due to
the economic opportunities provided by some of the milk nutrients contained in whey
(Jelen, 2002). Productions of whey proteins by ultrafiltration, lactose hydrolysis products,
and the use of whole whey or whey permeate as a fermentation feedstock are possible
options (Marwaha and Kennedy, 1988).

A large number of workers have carried out studies on the composition and
processing of whey for its use in foods and animal feeds besides studying the nutritive,
therapeutic and functional properties of whey. An overview of some of the recent works on
the above aspects is presented in this section.

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 Review of Literature

2.2.1 PRODUCTION OF WHEY

Currently, total world production of whey is approximately 85 million tonnes in
which India contributes approximately 8 million tonnes of the total global production
(Shukla et al., 2004; Raju et al., 2005; Misra, 2008).

Whey is good tasting, versatile and highly functional ingredient and possesses
benefits to food and beverage manufacture as they create notorious and delicious products
as per the demand of the consumers. The father of medicine, Hippocrates is said to have
insisted on a daily dose of two liters of whey. In the beginning of 17 th century, whey was
used to treat jaundice infected lesions of skin, gonorrhea and epilepsy (Gandhi and Dixit,
2008).

In India there has been a substantial increase in the production of Paneer in organised
sector resulting in increased availability of whey. It is estimated that about 2% of milk
produced in India is converted to Paneer and Chhana (Aneja et al., 2002) and production of
whey due to this is estimated at around 270 million Kgs per annum which contain about
16.2 million Kgs of valuable milk solids. Even though, whey contains valuable milk solids
it is hardly used for human consumption and largely wasted in large quantities adding to
environmental pollution (Jindal et al., 2004).
Many attempts have been done on utilization of whey in the formulation of various dairy
products but, still there is a lot of scope to explore the possibilities for its utilization in
beverage particularly in health based energy drink production industries (Singh et al.,
2009). The utilization of whey as a beverage production is more economical and value
addition than other methods of treatment and disposal.

2.2.2 WHEY AND ITS BY-PRODUCTS

Whey is a dilute, highly perishable greenish yellow fluid and the largest by-product of the
dairy world produced during the manufacture of cheese, casein, chhana, paneer, chakka
and coprecipitates. Its chemical composition and acidity varies widely (Table 1). It, in
general, contains about half of the total solids of milk, and is a source of precious nutrients
5
 Review of Literature

like lactose, whey proteins, minerals and vitamins. Whey proteins, though present in small
quantities, have high protein efficiency ratio (3.6), biological value (104) and net protein
utilization (95), and are next only to egg protein in terms of nutritive value (Renner, 1991).
Further, being a rich source of lactose, whey is good fermentation media for a number of
fermented products. In many applications, lactose in whole or deproteinised whey is
hydrolysed to glucose and galactose, thereby increasing its sweetness. Such lactose
hydrolysed syrups, generally after condensing, are mostly utilized in sweet confectionary
products and ice-cream. The market for products is estimated about $ 6.5 billion in sales
globally. Future growth is expected to be led by the industry’s increasing focus on
nutritional products, particularly in the dietary, sports and clinical segments of the market.

Table 1. Composition of Different Whey Systems (Monograph of IDA, 2007)

Constituents Cheddar Acid Rennet Chhana and Coprecipitates

cheese Casein Casein Paneer

Total solids (%) 6.7 6.9 6.7 6.4 6.1

Fat (%) 0.3 0.1 0.1 0.5 0.1

Protein (%) 0.9 1.0 1.0 0.4 0.3

Lactose (%) 4.9 5.1 5.1 5.0 5.1

Ash (%) 0.6 0.7 0.5 0.5 0.6

T. A (%) 0.2 0.4 0.2 0.4 0.3

The continuing annul growth in the production and consumption of cheese and
coagulated milk products represents the generation of extremely large additional quantities
of whey because of the huge base. The current world production of whey is estimated at
about 165 million tonnes. Cheese whey accounts for nearly 95 % of total whey. In India,
the major source of whey is from the production of chhana and paneer. In the absence of
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 Review of Literature

systematic surveys/statistics, the predicted value for whey production is estimated at 5

million tonnes per annum. In view of low solids contents of whey, there has been a gross
lack of interest in its utilization compared to other fluid by-products of dairy industry.

Utilization of whey has been of a great concern in the dairy industries engaged in
manufacture of cheese and coagulated milk products. The techno-economic problems
associated with the utilization of whey have been receiving consideration attention and
remarkable advancement have been made. For more than 25 years, a virtual explosion f
papers and reports has appeared about whey and its by-products, by and for scientists in
industry and academia. Whey proteins, together with lactose, have provided an excellent
area for research. Today, modern industrial processing techniques such as ultrafiltration
(UF), reverse osmosis (RO), new drying methods, hydrolysis, electrodialysis, ion-
exchange, fermentation and protein fractionation, among others, have converted whey into
a major source of ingredients with differing functional and nutritional properties, that could
be used in various branches of food and dairy industry. The global market for whey
derivatives is growing at an average of around 10 % annually. The predominant driving
force behind the development of whey utilization has been stringent regulation imposed by
the environment pollution agencies all over the world. Other aspect relates to economic
return from whey, which contains almost half of the solids of original milk. Food
manufactures are increasingly viewing whey products as an ideal means of achieving
added value.

Despite significant gains in the amount of whey being processed, a large amount of current
whey production still is disposed of as raw whey. Much of this represents production of
small plants, where the cost of purchasing, processing as well the subsequent transportation
and handling clearly exceeds the value of any whey products that might be produced. In
small plants, the choice remains some form of disposal, be its municipal treatment,
spreading raw whey on local farm lands for its nutrient value or feeding to local livestock.
Further, acid whey, because of high mineral content and low pH pose considerable
difficulties in utilization and therefore, mostly remain unutilized (Gupta, 1989).

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 Review of Literature

2.2.3. COMPOSITION OF DIFFERENT TYPES OF WHEY

The composition of different types of whey is tabulated below.
Table 2. Composition of whey (Khamrui and Rajorhia, 1998).

Sweet whey Medium acid whey Acid whey

Composition CM BM Paneer CM BM Casein Shrikhand

cheddar cheddar whey chhana chhana whey whey
cheese cheese whey whey (HCl)
whey whey
Solids % 6.35 6.87 6.06 6.25 6.40 5.95 6.96
pH 6.10 6.40 5.60 5.50 5.70 4.60 NA
Lactose % 5.00 5.01 5.03 4.86 4.90 4.30 NA
Protein % 0.90 0.98 0.30 0.72 0.89 0.70 0.38
Fat % 0.06 0.34 0.13 0.10 0.10 NA 0.43
Ash % 0.59 0.54 0.60 0.57 0.44 0.72 NA
Lactic acid 0.13 0.14 0.21 0.19 0.20 0.25 0.24
%
Calcium 448 501.5 710.65 526 446.3 1,298 NA
ppm

Phosphorus 486 441.5 560.5 493.7 533.3 1,770 NA

ppm

CM : Cow milk
BM : Buffalo milk
NA : Not available
Shashidhar, (2007) reported that paneer whey had an pH 5.32, acidity 0.22 per cent LA,
specific gravity 1.024, viscosity 1.068 (cP), water activity 0.995, total solids 6.21 per cent,
protein 0.39 per cent, fat 0.15 per cent, lactose 5.07 per cent and ash 0.60 per cent.

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 Review of Literature

In India there has been a substantial increase in the production of Paneer in organised
sector resulting in increased availability of whey. It is estimated that about 2% of milk
produced in India is converted to Paneer and Chhana (Aneja et al., 2002) and production
of whey due to this is estimated at around 270 million Kgs per annum which contain about
16.2 million Kgs of valuable milk solids. Even though, whey contains valuable milk solids
it is hardly used for human consumption and largely wasted in large quantities adding to
environmental pollution (Jindal et al., 2004).

2.2.4 SIGNIFICANCE OF WHEY

In the production of cheese, paneer and chhana, only 10-20 per cent of milk is
recovered as the desired end product and the remaining 80-90 per cent is whey. The paneer
and chhana contributes about 80 per cent of the total whey production (Shendurse et al.,
2009). Whey contains half of the nutritive milk solids. Each constituent of whey have their
own importance because of their nutritive, therapeutic and functional properties. Its
composition varies according to the type of product from which it is derived (Riedel,
1992).

Milk components not only provide nutritional security but also are capable of providing
potential health benefits in various forms. Many dairy ingredients are finding large-scale
application in nutraceutical products, which are possibly the hottest trend in the food
industry. Some dairy ingredients viz. whey carbohydrates and whey proteins are also
considered good prebiotics which affect the host by selectively stimulating the growth and
activity of one or more naturally present or administered bacterial species in the colon. The
energy value of whey is around 210 Kcal/lit (Swaminathan, 1982).

Whey nutrients represent a huge quantity of nutritionally rich food. The most valuable
components of whey are whey proteins. The most valuable components of whey are whey
proteins. Whey proteins have been regarded as superior to most of the other proteins such

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 Review of Literature

as egg, beef, casein and soya proteins in nutritive values. The amino acids profile of whey
protein shows that they contain all essential amino acids in excess of FAO standards. They
contain 11.3, 2.4, 2.8, 8.4, 11.8, 7.6, 3.6, 2.4 and 7.2g/100g protein of lysine, methionine,
cystine, threonine, leucine, isoleucine, phenylalanine, tryptophan, and valine respectively.
Biological value of whey protein is higher (104) as against whole egg (100), rice (74),
soya (59), wheat proteins (54) and casein (77). They also have higher Protein Efficiency
Ratio (PER) and Net Protein Utilization (NPU than casein. The PER of whey protein is 3.6
as against 3.8 for egg protein and 2.9 for casein, whereas NPU is 95 for whey protein as
against 93 for whole egg and 76 for casein (Renner and Abd-El-Salam, 1991).

In view of excellent nutrient and biological value, whey being used since 460 B.C for
treating an assortment of ailments. In middle ages, whey was recommended by many
th
doctors for treating varied diseases and by mid 19 century, whey cures reached a high
point with the establishment of over 400 whey houses in Western Europe (Poonam, 2007).

In Central Europe dyspepsia, uremia, arthritis, gout, liver disease, anemia and
even tuberculosis were treated with the whey injection (Holsinger et al., 1974). Delaney
(1976), has reported that whey protein concentrates are receiving considerable attention as
base material for preparation of a range of dietetic/therapeutic products, e.g. electro
dialysed whey based foods for use in the treatment of chronic uremia, application of whey
protein concentrate in a number of clinical disorders, use of whey protein concentrates and
whey protein fractions in the manufacture of non-residue preparations designed for
prophylactic cardiac dietary regimes.

Shahani et al. (1978) reported that whey and whey protein concentrates could
be extensively used for a number of formulations in human foods because of their high
nutritive value and especially for their therapeutic value. Whey could be used in infant
food formulae e.g. whey protein concentrates and specialized diets for invalids, geriatrics
and infants suffering from cardiac malfunction and convalescent patients.

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 Review of Literature

Lactose, often referred to as milk sugar, is the primary milk constituent in whey
which contributes around 80-90% of whey solids. Lactose is commercially available in
different qualities, of which major distinguishable categories are food-grade and
pharmaceutical-grade. The food sector remains the largest user of lactose, where in many
products it is partially a functional replacer of more expensive dairy ingredients and it is
also widely used in infant foods. Its specific functional properties have made lactose as
preferred choice of the pharmaceutical industry as a filling substance for pills and capsules
as well as an inert carrier for inhalation medicines (Zadow, 1991).

The major mineral components in liquid whey contain the mineral cations like
sodium, potassium, calcium and magnesium as well as anions like chlorine, citrate and
phosphate (IDF bulletin, 2003). About 40% of calcium and 43% of phosphorus in milk are
found in whey derived during Cheddar cheese making. Calcium has become the most well
known mineral available in milk since consumers became aware of its benefits in
preventing osteoporosis. Sodium and chloride constitutes the major portion of total ash in
whey (Khamrui and Rajorhia, 1998).

Considering the nutritional and functional virtues of whey, several attempts have
been made earlier to utilize it in bakery products, soups, beverages, specialized processed
foods, infant foods, sports foods, geriatric foods, chocolate and confectionaries, jellies and
custards, rice, chapatti, idly, dosa and low fat foods. However, keeping in view the
quantum of whey production and the problems associated with its disposal, many more
efforts should be made to utilize this important by-product.

2.2.5 NUTRITIONAL QUALITY OF WHEY

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 Review of Literature

Whey contains precious nutrients like lactose, whey protein, minerals, and vitamins.
These nutrients have an indispensable value in human dietary requirement (Prendergast,
1985; Mathur et al., 1988).

Whey protein is nutritionally superior functional food supplements compared with

other dietary proteins (Vasiljevic and Shah, 2007). They are extensively used in various
food applications, with an increasing global demand. Fox and McSweeney, (2003), Whey
proteins are highly soluble over a broad range of pH, (Zhu and Damodaran, 1994), a
property that is important in their application as foaming, emulsifying, gelling, and water-
binding agents in various types of food products. Whey proteins are used in sport
beverages, liquid meat replacements, baked products, processed meats, salad dressings, ice
creams, artificial coffee creams, soups, and various dairy products (Fox and McSweeney,
2003; Fitzsimons et al., 2007).

The most valuable component of whey is the protein, which is most effective in
meeting the body’s energy and amino-acid requirements, and is more superior to the
proteins available from other sources (Werner, 1981; Belem and Lee., 1999). The
individual components of whey proteins such as α-Lactalbumin is used in humanized
infant food formulations, whereas, β-Lactoglobulin is used in sports and dietetic beverages
(Durham et al., 1997).

About 3 million tonnes of whey is produced annually in India containing about 2 lakh
tonnes of valuable milk nutrients (Naik et al., 2009). The proteins present in whey
comprise of about 50 per cent beta-lactoglobulin, 25 per cent alpha lactoalbumin and 255
other proteins with a balanced spectrum of amino acids (Misra, 2008).

A compromised immune system can benefit from a diet rich in whey proteins. A
powerful anti-oxidant, glutathione, has the ability to enhance the body’s immune system.
Whey proteins are rich in sulfur-containing amino acids (e.g., cysteine, methionine) which

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 Review of Literature

are rate-limiting for the biosynthesis of glutathione, an antioxidant, anticarcinogen, and

immune stimulating sulfur containing tripeptide (German et al., 2001).

Consumption of the whey can supplement much of the lost organic and inorganic
nutrients to the extra cellular fluid and utilization of these fluids can be targeted to the
people working with strenuous occupation like sportsman, body builders, competitive
athletes, exercising for pleasure and other people with similar kind of activity (Prendergast,
1985; Singh et al., 2009).

The paneer whey contains sodium 350 mg/l, potassium 1300 mg/l, calcium 480 mg/l,
magnesium 59 mg/l, chloride 1349 mg/l, citrate 6750 mg/l, zinc 280 μg/l, total protein 0.41
per cent, fat 0.01 per cent, lactose 4.5 per cent, total solids 5.8 per cent, pH 5.5 (Nupur and
Gandhi, 2009).

2.2.6 Therapeutic benefits

Holsinger et al., (1974) reported that whey has prophylactic effect against
tuberculosis and arthritis. Whey and whey based products have relatively high lactose
content, which forms a suitable substrate for probiotics in the intestine (Renner,
1992).whey is also helpful in curing gastrointestinal disorders, anaemia and liver problems
(Jelen, 1992).

Whey proteins are generally regarded as safe for food applications

(Hugunin, 1987; Morr, 1987) because of their excellent nutritional and functional
properties. Gupta (2000) reported that whey proteins are next to egg protein in terms of
nutritive value. WPC has been considered as a significantly nutritional ingredient to
improve nutritional profile of varieties of foods as recorded by many workers (Jelen, 2002;
Huffman, 1996).

Delaney (1976) reported that whey protein concentrates are receiving

considerable attention as base material for preparation of a range of dietetic/ therapeutic

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 Review of Literature

products, such as electrodialysed whey based foods for use in the treatment of chronic
uraemia, whey protein concentrate for a number of clinical disorders, whey protein
concentrates and whey protein fractions in the manufacture of non-residue preparations
designed for prophylactic cardiac dietary regimes.

In Central Europe, dyspepsia, uraemia, arthritis, gout, liver disease, anaemia and
even tuberculosis were treated with the injection of up to 1500 g of whey per day
(Holsinger et al., 1974).

2.2.7 Functional properties

Apart from their nutritional and therapeutic advantages, whey proteins exhibit
excellent properties such as solubility, foaming, emulsifying, gelling and water binding etc.
(Methews, 1984; Patel and Kilara, 1990). Whey protein is commonly marketed and
ingested as a dietary supplement, and various health claims have been attributed to it in the
alternative medicine community (Marshall, 2004). Whey proteins contain more essential
amino acids and branched-chain amino acids than most other food proteins, and as a
consequence are associated with the modulation of insulin responses in humans (Etzel,
2004; Pfeuffer and Schrezenmeir, 2007).

Whey shows a wide range of functional characteristics contributing to

flavour, colour, texture and overall acceptability attributes in various varieties of foods
(Marshall, 1995; Regester et al., 1996). Additionally branched-chain amino acids from
whey including isoleucine, leucine, and valine have been linked with postprandial
stimulation of insulin and increased plasma amino acid levels (Matthews, 2005).

Functional properties of proteins are those physico-chemical properties which

govern the performance and behavior of proteins in food systems during their preparation,
processing, storage, and consumption, i.e., properties affecting final quality of foods
(Kinsella and Whitehead, 1989).

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 Review of Literature

Several workers (Townsend and Nakai, 1983; Shimizu et al., 1985; Kato et al.,
1985) stated that the segmental molecular flexibility may predominate structural factor in
determining the foaming capacity and attributed that the surface hydrophobicity may be the
most important factor, which affects emulsifying activity. Some of the applications
wherein the various functional properties of whey proteins are utilized are listed below.

Table 2.3 Functional properties of whey proteins in various foods (Cheftal et al., 1985).

Food Desirable functionality

Beverages Solubility at wide pH range, heat stability,

viscosity.
Soups, Sauces Viscosity, emulsion, water retention
Dough formation in Bakery products Formation of matrix by film with viscoelastic
(e.g. bread and cake) properties, cohesion, heat denaturation, gelation,
water absorption, emulsification, foaming and
browning.
Dairy products (e.g. processed Emulsification, fat retention, viscosity foaming,
cheese, ice-cream, desserts) gelation and coagulation.

Meat products (e.g. sausages) Emulsion, gelation, water and fat absorption and
retention.
Egg substitutes Foaming, gelation.
Meat extenders (e.g. Texturized Water and fat absorption and retention,
vegetable protein) insolubility, hardness, cohesion and heat
denaturation.
Food coating Cohesion, adhesion.
Confectionary products (e.g. Milk Dispersibility, emulsification.
chocolate)

2.3 Environmental considerations

Whey have a BOD in the range of 3.50x104 to 4.00 x104 mg/l (Marwaha and
Kennedy, 2007; Anekar and Rao, 2009). Due to the higher BOD of whey, it poses a major
world-wide disposal and pollution problem; thus an effective and permanent solution is
urgently needed (Venkatraman and Achi, 2004).

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 Review of Literature

About 40per cent of the total global production of whey is disposed as raw whey
(Reddy et al., 1987) causing serious problems of environmental pollution due to high
organic matter content. It has been estimated by the Water Pollution Research Laboratory
that whey has a BOD of 38,000 to 46,000 ppm, even in some cases it reaches up to 76000
ppm as compared to 200 ppm permissible limit for domestic sewage (NDRI, 1978; Misra,
2008). Whey causes an environmental pollution, which is being considered one of the most
polluting food by/co-product streams ((BOD) >35,000 ppm; (COD) >60,000 ppm) (Siso,
1996). The Government of India has promulgated the Environmental Protection Act-1986
that makes it obligatory to pre-treat whey before discharge in inland water or rivers.
A large portion of whey from the organized dairy sector is not being utilized
and being disposed through the effluent treatment systems. Older methods of disposal are
becoming less available and several other possible methods are prohibitively expensive to
operate (Smithers, 2008). Whey disposal is a serious problem for dairy industry. In order to
reduce pollution load, whey should be treated to obtain commercial products (Gupte and
Nair, 2010). In recent times, the environmental regulations have become stringent across
the world necessitating the treatment of whey prior to disposal through sewage system. A
recent study indicates that treating 5 lakh liters of whey in the sewage would cost $ 10,000
per day for primary treatment or $ 145,000 for tertiary treatment (Khamrui and Rajorhia,
1998). In India it is estimated that the cost of treatment of each litre of Paneer whey is
around 50-60 paise which is considered expensive.
Because of problems involved with the whey disposal, since long time efforts are
being made to utilize whey for beneficial purpose and the benefits of utilization of whey
solids is reviewed.

2.3.1 UTILIZATION OF WHEY

Whey and whey derivatives have been utilized in a number of other products such as
snacks, dairy, meat products, beverages etc. They are also used in the production of frozen
desserts, ice-creams, confectionery, salad dressings, and sport supplements (Aguilera,

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 Review of Literature

1995). Whey proteins have wide range of applications in the processing of foods such as
bakery products, dairy products and beverages. Some of the salient works are listed:

Table 2.4 Utilization of whey in foods and their functional benefits

Product Form of whey utilization Functional benefits

Snacks and Whey powder, WPC, WPI, Skim milk powder and sucrose substitute,
confectionery (e.g. lactose hydrolyzed whey, improvement in body and color, improved
milk chocolate, whole whey, demineralised water binding, enhance flavor, gelling and
caramels, butter and delactosed whey and emulsifying agent, stabilize coating on
scotch etc.) condensed whey. confectionary, lactose assists crystal
formation in butter scotch, confectionary
remains fresher for long .

Dairy products Whole whey, dried whey, MSNF substitute enhanced sensory,
(e.g.processed WPC, concentrated whey, rheological and nutritional profile as a result
cheese, ice-cream, condensed whey. of emulsification, fat retention, viscosity,
yoghurt, desserts) foaming, gelation and coagulation.
Meat products WPC, Dried whey, reduced Binder, tenderizer, emulsion stabilizers,
lactose whey, reduced gelation, water and fat absorption and
minerals whey, fermented retention.
rennet whey.
Infant and weaning Whey powder, Improved reconstitution properties, imitation
foods Demineralised whey human milk.
powder.
Beverages (e.g. fruit Whole whey, WPC, Solubility at wide pH range, heat stability,
flavored, carbonated, whey powder, viscosity, enhanced nutrition.
high protein, deproteinated whey,
alcoholic, meal fermented whey.
replacement, sports
nutrition, etc.)

Dietetic foods (e.g. Demineralised and Caters to special nutritional

geriatric foods) delactosed whey demands for high protein, vitamins
powder, whey protein and minerals, enables easier and
hydrolysates, WPC, quick digestion
whole whey, whey powder.
Soups and gravies Dry whey, WPC, WPI, Viscosity, emulsion, water retention,
demineralised and desirable body and flavor.
delactosed whey powder.

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 Review of Literature

2.3.2 BENEFITS OF WHEY UTILIZATION

Utilization of whey results in reducing the wastage of valuable nutrients which

can be diverted for the beneficial use in the form of food. The various nutrients present in
whey can supplement the energy value of a variety of foods consumed by the lower
income group of population across the world. A variety of functional foods, geriatric and
pediatric products are produced by beneficially utilising the valuable proteins and minerals
constituents present in whey (Puranik, 2000). Whey can effectively be used as one of the
ingredients in the animal feed ration (Schingoethe, 1976) and the effective utilization will
considerably reduce the pollution for the Cheese and Paneer factories. Thus utilization of
the whey solids is in the larger interest of the society and thus attempts are continuing for
developing appropriate processes for using the whey solids beneficially.

2.3.3 UTILIZATION OF WHEY IN LACTOSE PRODUCTION

Lactose and lactose-derived products are playing an increasingly important role in

various branches of food industry. Dry matter of whey is about 6 to 7 %, of which lactose
is about 75 % (Gonzalez siso, 1996). The disposal of whey is a great environmental
problem because of its very high Biological oxygen demand (BOD). Since, lactose alone is
responsible for 80 % of BOD (Mawson, 1994), attempts have been made by different
researchers to solve this problem by isolation of lactose or by its conversion to single cell
protein, lactic acid, ethanol and methane. Lactose has been isolated from whey by four
different procedures: a) conventional concentration procedure, b) reverse osmosis and
ultra-filtration, c) precipitation with alkaline earth metal chlorides and d) precipitation by
different alcohols.
The yield, i.e. percent of lactose isolated varies from 60.37 to 90.27 %, depending
upon the procedure adopted. Best quality lactose with 99.99 % purity was obtained by
ethanol extraction preceded by deproteinization and demineralization of whey and it
contains only 0.04 % ash and protein and fat are below detectable limit, which makes this
method the most suitable for use in pharmaceutical industries (Paul et al., 2002). Lactose
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 Review of Literature

has various applications in food and pharmaceutical industries due to its multiple
functional properties (Kapil et al., 1991).

2.3.4 UTILIZATION OF WHEY FOR HUMAN FOODS

In order to utilize the nutrient rich whey several attempts have been made
worldwide and some of the products so developed are commercially available in the
market. Some of the whey solids based foods with their specific role are summarized in
table 2.3.

Table 2.3: Commercially available whey foods: - (Vyas et al., 1980)

Food %Whey Contribution

solids
used
Dried infant food 25-40 Nutritional, biological
Beverages 6 Flavour, body
Dried soups and gravy 50-75 Flavour, body
Bases
Dried culture media 85-97 Nutritional
Bakery products 3-10 Flavour, texture, keeping quality
Confectionary products 4-10 Flavour, moisture, whipping property
Frozen desserts 3-4 Flavour, fruit stability
Cheese foods 10 Flavour, body
Dry mixes 10 Tenderizing, colour carrier for fats and
Oils

2.4 LACTOSE
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 Review of Literature

Lactose is carbohydrate of milk and is the only sugar of animal origin. Lactose in its
pure form is a white, water-soluble crystalline powder of moderately sweet taste with no
odour. Lactose, a disaccharide that comprises the monosaccharides glucose and galactose,
is the primary carbohydrate found exclusively in mammalian milk. Absorption of lactose
requires lactase activity in the small intestinal brush border to split the bond linking the
two monosaccharides.

Whey contains approximately 6 per cent total solids of which 70 per cent is lactose
and 11 per cent is whey protein (Zall, 1984). Paneer whey contains 5.03 per cent lactose
(Khamrui and Rajorhia, 1998).

2.4.1 LACTOSE INTOLERANCE

Lactose intolerance is a frequent clinical syndrome causing non-specific

gastrointestinal symptoms such as flatulence, abdominal pain excessive passing of gas and
diarrhoea (Marek et al., 2007).
Lactose intolerance refers to gastrointestinal symptoms experienced by some
individuals who have low levels of lactase. Individuals with lactose intolerance are unable
to digest significant amounts of lactose due to an inadequate amount of the enzyme lactase
(Swagerty et al., 2002).
Lactose is the major carbohydrate in milk and some other dairy foods. If the
activity of the lactase enzyme is low, undigested lactose may reach the large intestine
where it is fermented by naturally residing gas-producing bacteria. This can lead to
symptoms of lactose intolerance.

2.4.2 LACTOSE HYDROLYSIS

In the lactose hydrolysis process, the disaccharide lactose is converted to its

monosaccaride units, β-D glucose and β-D galactose (Puranik and Kanawjia, 1998). The
conversion of lactose to monosaccharides results in more sweetness, solubility, easily
fermentable and directly absorbed from the mammalian intestine. There are many methods

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 Review of Literature

of methods available for lactose hydrolysis. Acid hydrolysis includes direct acidification
and ion exchange hydrolysis; now-a-days cation exchange resin is preferred. For the
hydrolysis of pure lactose or lactose in milk and whey products enzymatic hydrolysis
prevails in food technology (Ryder, 1988). Many research efforts have been focused on
ways to reduce or remove the lactose in dairy products (Kocak and Zadow, 1989).
Lactose-hydrolysed products are good for the lactose intolerance and easy digestion
aspects. However, this lactase has additional health benefits. An example is a sweeter taste
without extra calories. Glucose is much sweeter than lactose. This means that lactose-free
products are also naturally sweeter - without the use of sugar or artificial sweeteners.
This enzymatic lactose hydrolysis process offers alternative ways for the utilization of
whey. Hydrolysis of lactose into simpler sugars i.e. glucose and galactose improves
desirability (Geilman, 1993) by increased flavour enhancement, increased osmotic
pressure, easier digestibility, increased solubility, increased sweetness and easier
fermentability (Coton, 1980; Arndt and Wehling, 1989; and Timmermans, 1997).
In comparison to sweetness of sucrose (taken as a base with a relative value of 100)
relative sweetness of lactose, glucose and galactose are 16, 74 and 32 %, respectively
(Pazur, 1970). Relative sweetness of lactose is approximately one-sixth that of sucrose
(Fennema, 1996). It has been reported that the sweetness of a hydrolyzed lactose solution
could be 50 % higher than the un-hydrolyzed solution (Zadow, 1991). According to Zadow
(1986), hydrolysis of 70% of the lactose in milk increases its sweetness by an amount
comparable to the addition of approximately 2% sucrose. It has been reported by many
workers that hydrolysis increases the sweetness (Coton, 1980; Arndt and Wehling, 1989;
Geilman, 1993; and Timmermans, 1997) which is due to release of glucose and galactose,
and which are known to be sweeter than lactose. An increased sweetness of lactose
hydrolyzed whey products provides commercial opportunities for whey utilization.
β Galactosidase is widely used in food industry to improve sweetness, solubility, flavor
and digestibility of dairy products This enzyme has been used as a model for studying its
activity in amorphous matrices (Burin and Buera, 2002).

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 Review of Literature

Enzymatic hydrolysis of lactose by β galactosidase is one of the most popular

technologies to produce lactose reduced milk and related dairy products for consumption
by lactose intolerant people (Ladero et al., 2000; Ladero et al., 2005). Lactose is a
hygroscopic sugar and has a strong tendency to absorb flavors and odors and causes many
defects in refrigerated products such as crystallization in dairy foods, development of
sandy or gritty texture and deposit formation (Panesar et al., 2007). By hydrolyzing lactose
with β galactosidase, the problems associated with whey disposal, lactose crystallization in
frozen concentrated deserts and milk consumption by lactose-intolerant individuals can be
eliminated.

The intestinal deficiency of β-galactosidase some in population of some countries

shows lactose intolerance and they have difficulty in consuming milk and dairy products
(Vassilis and Miguel, 1985; Suarez and Savaino, 1997). The prevalence of lactose mal-
absorption is high in East and South Indian adults (Johnson, 1981). Enzymatic hydrolysis
of lactose by β galactosidase is one of the most popular technologies to produce lactose
reduced milk and related dairy products for consumption by lactose intolerant people
(Ladero et al., 2000; Ladero et al., 2005)

2.4.3 APPLICATION OF LACTOSE HYDROLYZED WHEY IN DAIRY

AND OTHERS FOODS

Hydrolysis of the lactose reduces protein precipitation in drinks and avoids the lactose
intolerance problem (Dahlen and Strining, 1984). Bernal and Jelen (1989) studied the
effectiveness of lactose hydrolysis in cottage cheese whey based drink using six
commercial lactase preparations (two neutral enzymes from Kluyveromyces marxianus
var. lactis and Kluyveromyces fragilis and four acid enzymes from Aspergillus oryzae).
The hydrolysis conditions for neutral enzymes were 38⁰C and pH 6.8 and for acid enzymes
at 55⁰C and pH 4.65, which was giving 80 % hydrolysis.
Lactose hydrolysed whey prevents moisture loss from caramels during storage
because of its humectants property (Guy, 1978). Haven & van Gennip (1983) report the

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 Review of Literature

following advantages while using hydrolysed whey at levels of 50% and 100%
replacement of the skimmed sweetened condensed milk:
1. The caramels were darker in colour and higher in flavour.
2. Caramels were chewier due to the higher levels of whey proteins.
3. Crystallization of lactose did not occur due to the inhibiting effect of the glucose
and galactose.

4. Increase the yield of final product by adding whey solids to the products.

A Swedish Dairy Co-operative Arla Group hydrolyzed the whey permeate and
used for preparation of soft drink (Anon, 1983). Lactose hydrolyzed permeate beverage
was prepared by the addition of Lactozyme (Kluyveromyces lactis) with an activity 3000
LAU / ml at rate of 1ml/ lit at 37⁰C at pH 6.5 (Suresha and Jayaprakasha, 2004). Paul
(1990) hydrolyzed the lactose in whey by adding 0.25-1.0 g / lit of lactase enzyme. A
fermented drink was made by hydrolyzing the whey using Maxilact at the rate of 0.3 g / lit,
which resulted in 80 % hydrolysis of total lactose (5.1 %) in 30 min (Kar et al., 1998). In
Sweden, a drink which is a mixture of tropical fruit juices, hydrolyzed lactose and whey
protein concentrate has been introduced. The product has recently been launched in the UK
under the name of “Nature's Wonder”. It is understood that a similar product is also
available in Germany. The drink thus uses the sweetening power of the glucose and
galactose but at the same time gains the advantage of having the presence of whey protein
which have a high nutritional value.

Hydrolyzed whey syrup could also be used to replace a part of the milk powder
in the recipes. It is suggested that up to 50% replacement could be achieved (IDF, 2003).
Drobot et al, (1983) used cheese whey with 50% of its lactose partially hydrolyzed in
bread formulations at 20% flour weight and the resultant bread obtained had good volume,
better porosity and better compressibility with good keeping quality and lasting freshness.

Whey solids are used in different confections such as toffees, caramels and fudge, largely
for the beneficial effects of the whey protein on texture (Webb, 1966). Hydrolysis of
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 Review of Literature

lactose in whey helps the use higher level of whey solids without lactose crystallization
(Castelao et al., 1981).

2.4.4 APPILICATION OF HYDROLYZED LACTOSE IN PROCESSING

A report from the USA(Anon, 1982) describes a process for hydrolysis of cheese whey,
using immobilized enzyme of fungal origin producing products combining the functional
properties of whey proteins with the sweetening power of a glucose/ galactose syrup.
These ingredients can be utilized in various products including ice cream.

Liquid milk: Lactose hydrolysis in liquid milk improves digestibility for lactose intolerant
consumers. In flavoured milks, lactose hydrolysis increases sweetness and enhances
flavors (Shilpa, 2010).

Bread and bakery products: Use of hydrolysed lactose syrup (50, 75, 95%) from whey
permeate as sugar substitute in milk bread formulation with or without demineralization
were decolorized and concentrated to 50% solids to give hydrolysed syrup (HLS). These
were added to bread mix at 6% of the flour content. Bread made from 95% HLS was
comparable with control bread pH, volume, texture and flavour. Addition of 3.9- 4.1%
w/w of the syrup (on flour) significantly improved the physicochemical and organoleptic
properties of wheat bread. Hydrolyzed lactose whey is also used in the manufacture of
biscuits (Kanawjia, 1992).

Frozen milk and milk shakes: The concentrated, decolorized, demineralized, lactose
hydrolyzed whey permeate were used successfully in mixes for soft-serve ice milk,
hardened ice milk, and milk shakes, but only small amounts of permeates could be
incorporated (Kanawjia, 1992).

Milk powders: Lactose hydrolysed milk powders for dietetic uses, especially for infants
with temporary lactase deficiency (Shilpa, 2010).

Fermented milk products: In some cases, lactose hydrolysis in milk used for the
manufacture of cheese and yoghurt can increase the rate of acid development and thus
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 Review of Literature

reduce processing time. Cheese maturation time may also be reduced, possibly through the
presence of small amounts of proteases in some lactase preparations. Yoghurt produced
from lactose hydrolyzed is highly popular and is targeted to the health conscious
consumers (Kanawjia, 1992).

The use of lactose hydrolysed whey in yoghurt (Whalen et.al., 1988) and whey beverages
(Dwivedi.et.al., 2006) have also been reported.

Concentrated milk products: Lactose hydrolysis in concentrated milk products (eg.

sweetened condensed milk, or cream), prevents crystallisation of lactose (Shilpa, 2010).

OTHER APPLICATION OF HYDROLYZED WHEY

(a) Deproteinised whey (permeate): After demineralization, lactose hydrolysed permeate

is evaporated to syrup containing 60-65 percent solids. This has properties very similar
to those of medium dextrose equivalent to glucose syrup.

(b) Whey for animal feed: Lactose hydrolysis in whey enables more whey solids to be
fed to pigs and cattle and also prevents crystallisation in whey concentrate.

(c) Whey: Lactose hydrolysed whey is concentrated to produce syrup containing 70-75
percent solids. This syrup provides a source of functional whey protein and sweet
carbohydrate and is used as a food ingredient in ice creams, bakery and confectionary
products.

Use of lactose in nutritious products is limited because of low solubility, low sweetener
power and laxative effect, if consumed in high concentration. The hydrolysis of lactose in
glucose and galactose by b-galactosidase is one important process in the food industry, due
to the potentially beneficial effects on assimilating the foods containing lactose, as well as
the technological and environmental advantages of industrial applications (Marcela et.al.
2008)
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 Review of Literature

1. Production of milk with sweetened flavor & of good acceptability, that can be
consumed by people with intolerance to this sugar.
2. Formation of galacto-oligosaccharides during lactose hydrolysis to favor the
growth of intestinal bacterial microflora.
3. Improvement in the technological and sensorial characteristics of foods containing
hydrolyzed lactose from milk or whey, such as increased solubility (avoidance of
lactose crystallization and the grainy aspect of ice-cream and condensed or
powdered products) and higher sweetening power (and thus lower caloric content
of the products).
4. Better biodegradability of whey after lactose hydrolysis (Zuzana, 2006).

2.5 FERMENTATION
Fermentation is one of the oldest forms of food preservation and biological
upgrading of dairy by-product into value added food has been well established. Whey can
be used in the manufacture of fermented products e.g. acidic and alcoholic beverages,
yeast, alcohol, lactic acid, vitamins and vinegar etc. Being a rich source of lactose, whey is
a good fermentation medium for a number of fermented products.

The disposal of whey may be more problematic in future

because of the stringent anti-pollution regulations coupled with the growing consciousness
of people against environmental pollution. The only option open to the majority of the
manufacturers appears to be to use the technique of lactose hydrolysis. But it is capital
intensive and therefore unsuitable for small producers of chhana/paneer whey. The only
alternate available to them is the production of whey beverages by fermentation technique
(Misra, 2008).

2.5.1 FERMENTATION OF WHEY

Whey has been found to be useful as base for the growth of some of the LAB in
the preparation of beverages. These beverages are obtained due to lactic acid bacteria
fermenting lactose in whey to form mainly lactic acid. The lactic acid imparts fresh flavor
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 Review of Literature

and is known suppress the growth of pathogenic spoilage organisms. Antibacterial

substances produced during fermentation are reported to have curative properties in
controlling several gastro-intestinal disorders (Prasher and Gandhi, 1990).

An attempt has been made at National Dairy Research Institute at Karnal (India) to
convert whey into palatable refreshing probiotic in nature and low cost beverage named as
‘Acidowhey’ and its concentrate (Gandhi and Patel, 1992).

Drissen and Berg, (1990) emphasized the significant feature of adopting the process
of manufacturing of fermented whey beverages by dairy industries for the advantages.

2.5.2 FRUIT FLAVOURED WHEY BEVERAGES

India has not only made great progress in milk production, but it has also emerged
as the top fruit producer in the world (FAO, 1995). However, as much as 25-30 per cent of
the total fruit produced in India gets spoiled in the absence of infrastructure for appropriate
post-harvest technology. Another distressing aspect is that, the whey, which contains 6-7
per cent of milk solids, is considered as a waste product because of its low concentration of
milk constituents (Sienkiewicz and Riedel, 1990). Hence, formulation of new products in
the form of nutritious and refreshing beverages by using suitable combination of whey and
fruit juices both in concentrated and dried forms would permit economic utilization of
whey and value addition to guarantee high income. In 1987, the Federal Republic of
Germany considered whey drinks as mixed whey products for the first time and defined it.

Farah and Bachmann, (1981) reported that fruit juice concentration up to 30-
40 per cent using orange juice have been reported to be acceptable for the preparation of
fruit-flavoured buttermilk beverages. Whey beverages have been recognized as a genuine
thirst quencher, light, refreshing, healthful and nutritious (Prendergast, 1985).
Jayaprakasha et al., (1986) found 10 per cent sugar, citric acid and orange flavour as
optimum level for whey fruit flavoured beverage. Reddy et al., (1987) reported 8per cent
lemon juice and 14 per cent sugar level for developing acceptable quality beverage. In

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 Review of Literature

general, the total solid level above 15 per cent, sugar level between 7-10 per cent and pH
below 4.5 is required for developing acceptable quality beverage. Beverage with 10 per
cent sugar and orange flavour received highest score.

Gagrani et al., (1987) developed a fruit-flavoured whey beverage with 0.5 per cent
acidity and 20o brix using mango, orange, pineapple and guava juices at 10, 15, 20 and 25
per cent of whey. Highest sensory scores were obtained for the products containing 10 per
cent orange juice, 15 per cent pineapple juice, 15 per cent mango juice and 25 per cent
guava juice. Among these, mango flavoured whey beverage received maximum score for
colour, sedimentation, turbidity and viscosity. Yalcin et al., (1994) observed that in
chhana whey beverage, increased concentration of mango juice (due to colour) into the
drink increased its acceptability.

Khamrui, (2000) used cheddar cheese whey for the development of ready to serve
Kinnow juice beverage. It was developed by mixing Kinnow juice concentrate (23 o Brix by
reverse osmosis) with concentrated fresh cheddar cheese whey (45 per cent total solids)
along with other ingredients (sugar 7 per cent, pectin 0.05 per cent and carboxy methyl
cellulose 0.15 per cent). This formulation when reconstituted with three parts of water
received maximum organoleptic acceptability.

Shukla et al., (2000) developed a ready to serve beverage from whey by the
addition of 10 per cent sugar and 30per cent litchi juice. Suresha, (2002) reported that
whey blended with orange juice in the ratio of 70:30 (whey: orange juice) was more
acceptable by the panel compare to other combinations and he also concluded that out of
three juices i.e., mango, orange and pineapple juice, the pineapple juice blended sample
was highly acceptable by the panel of judges. Djuric et al., (2004) studied the effect of
quality of whey fruit juice (orange, pear, peach and apple) and they concluded that peach-
whey beverage containing 6 per cent of dry matter and 2 per cent of sucrose as well as
having pH 3.6 was best. Whey based fruits beverages are more suitable for health as
compared to other drinks (Sarvana, 2005).

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The beverage prepared by utilizing chakka whey with 15 per cent pomegranate
juice, had secured the highest sensory score (8.81) and ranked as most acceptable product
followed by 10 per cent with 8.65 points sensory score. The percentage of protein total
sugar, ash and acidity (per cent LA) of the product increased with increase in the levels of
pomegranate juice (Babar et al., 2008).

Attempts were made to develop a soft beverage from paneer whey and guava pulp
which pasteurized at different temperatures and timings for estimating its shelf-life. In the
preparation of beverages the volume of guava pulp (25 per cent), sugar (10 per cent) and
paneer whey (65 per cent) were kept constant while the pasteurization temperatures and
timings were varied from 60°C-70°C for 15-35 minutes. Whey-guava beverages
pasteurized at 70°C for 35 minutes was found to be best in terms of sensory quality after
45 days and pH, acidity, protein, total sugars and reducing sugars found to be high than
that of the other samples (Divya and Archana, 2009).

Whey Based Watermelon Beverage (WBWB) prepared by bending watermelon juice

(15 per cent), sugar (7 per cent) and different concentration of Betel leaves distillate (0, 1,
2 and 3 per cent) into chhana whey (78-75 per cent). The prepared beverage has red colour,
highly acceptable taste and overall acceptability. The overall acceptability of beverage
improved with increase in betel leaves distillate up to 2 per cent. After in-bottle
sterilization and cooling, the beverage was stored at refrigerated temperature. The storage
study showed that there is an increasing trend in the TSS, acidity, and reducing sugar and a
decreasing trend in the pH and ascorbic acid but total sugar has non-significant effect
during storage. The sensory quality of fresh beverage containing 2 per cent betel leaves
distillate on the preparation as well as 30 days of storage were found to be highly
acceptable. Additions of 3 per cent betel leaves distillate do not improve the mouth-feel but
they extend the storage period of the product. This indicates that antimicrobial and anti-
oxidant properties in essential oil of betel leaf increase the storage stability of the beverage
(Naik et al., 2009).

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Sensory quality of paneer whey beverage prepared with kokum juice was studied
with a view to standardize the process of utilizing paneer whey. Three levels of kokum
juice added are 2.5, 3.5 and 4.5 per cent and two sugar levels 12 and 15 per cent with 0.1
per cent jeera powder were selected for manufacture of six types of paneer whey beverage
samples. Raspberry red colour was added to match the appearance with kokum sherbet and
enhance the acceptability of the product. Paneer whey beverage prepared from 2.5 per cent
kokum juice and 15 per cent sugar with 0.1 per cent jeera powder was found sensorily
most acceptable (Rupnar et al., 2009).

Mango pulp added treatments scored a high by the sensory panel. Mango flavoured
sweetened whey drink can be prepared by using 4, 5 and 6 per cent of mango pulp with
highest consumer acceptability as compared to the control. Whey contains about half of the
milk solids and addition of mango pulp in the preparation of whey drink increased the
deliciousness and nutritional value (Pandiyan et al., 2011).

Whey and juice of mango (Cv. Kesar) were utilized at various combinations
(70:30, 75:25 and 80:20) for preparation of nutritious ready-to-serve (RTS) beverages and
evaluated for various physico-chemicals and sensory attributes during storage. The study
revealed that the RTS beverage prepared with 70 per cent whey and 30 per cent mango
juice scored maximum for almost all sensorial quality attributes such as appearance, color,
flavor, taste and overall acceptability and also found highest in ascorbic acid content
(9.80mg/100g). A reducing trend was observed in total sugars and ascorbic acid and
increasing trend was observed in reducing sugars and acidity content during the storage of
beverage at refrigeration temperature over a period of 30 days. The beverage remained
unchanged with respect to TSS content along the storage period (Sakhale et al., 2012).

Whey based beverage prepared from pineapple and bottle gourd juices in
combination with edible extract of herbal medicinal plants like Mentha arvensis will not
have only excellent nutritional properties but will also posses therapeutic, prophylactic,
antibacterial and organoleptic properties (Baljeet et al., 2013).
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 Review of Literature

The study was done on development of a probiotic beverage using whey and
pineapple juice. Lactobacillus acidophilus was used as the probiotic organism. The level of
pineapple juice addition was optimized on the basis of sensory quality evaluation.
Fermentation time using 1 per cent inoculum of L. acidophilus was optimized on the basis
of sensory quality evaluation, growth and activity in terms of pH and acidity. The 65:35
blend ratio of whey and pineapple juice fermented for 5 hrs gave desirable results with
highest sensory scores for overall acceptability and a total viable count of more than 10⁶
cfu/ml (Shukla et al., 2013).

2.5.3 QUALITY CHARACTERISTICS OF WHEY BASED FRUIT BEVERAGES

Despite its relative simplicity, finding a successful flavour combination to mask
the unpleasant whey taste is difficult. Several workers have reported cheesiness or saltiness
as two most dominant flavours in processed whey beverages. The presence of residual
lipids also sometime pose flavour problem in whey beverages especially if the product is
thermally processed at elevated temperature. The whey flavour was observed to be most
compatible with citrus flavour (Holsinger et al., 1978). Branger et al., (1999) evaluated the
sensory characteristics of cottage cheese whey and grapefruit juice blends. They observed
that increasing the proportion of whey increased the cheesiness and saltiness in blends.
Deproteination had no effect on sensory attributed however vacuum stripping reduced
cheesiness with subsequent improvement in grapefruit flavour and perception of
sweetness. Consumer prefer whey beverages that are sparkling clear, hence clarification of
whey is essential to remove residual lipids, casein fines and partially denatured whey
proteins. Acid whey possesses fewer proteins as compared to sweet whey and that make
them more suitable base for beverages base. Major problems reported in whey beverages
are turbidity and sedimentation after heat treatment and during subsequent storage. During
thermal treatment whey proteins coagulate near their isoelectric point (at acidic pH) which
causes development of turbidity and subsequent sedimentation in whey-fruit juice beverage
(Gagrani et al., 1987). Shekilango et al., (1997) noted that the other major problems often
encountered in whey-fruit beverage are cloudiness and high viscosity.
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 Review of Literature

Sensory testing of beverage using consumer panel indicated that sourness and acidity
are critical quality parameters. The storage temperature had significant effect on
sedimentation and serum separation of beverage and higher storage temperature maximize
these defects. According to Jelen, (1992), such quality problems often lead to failure of
these products in market. Several approaches have been tried to minimise these problems.
Passing whey through a centrifugal classifies removed casein fines and residual fats and
improved the clarity (Jelen et al., 1987). Another approach adopted was to homogenize the
product after pasteurization (Barabas and Albrecht, 1988). According to them whey drinks
processed in this way were rated as homogenous with excellent flavour quality.

Beverages and soups based on whey continue to receive a considerable amount of

attention nowadays. These indicate the growing awareness among consumers and
manufacturers alike for the enormous potential these offered for diversifying product
profile. Technological packages are available for wide range of whey based such products
and with better understanding regarding the functionality of different ingredients it would
be possible to develop “functional” whey products for specific target groups (Ashish,
2008).

2.5.4 STABILIZER LEVELS

Mehanna and Mehanna, (1989), stated that the sensory score of whey drink
increased with level of stabilizer up to 0.35 per cent; use of 0.5 per cent resulted in
undesirable flavour. Sameen et al., (2013), reported that incorporation of stabilizer
exceeding 0.3 per cent adversely affected the sensory properties of dairy products. Boland
et al., (2006), reported that addition of hydrocolloids improved the texture of the whey
beverage, it influenced in a decline of perceived flavour intensity in the product.

2.5.5 CARBONATED BEVERAGES

The major mechanisms of spoilage of most dairy products are microbial, since milk
solids act as a good environment for their growth (Hotchkiss et al., 2006). The major

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 Review of Literature

spoilage organisms for dairy products include aerobic psychrotrophic Gram-negative

bacteria, yeasts, moulds, heterofermentative lactobacilli, and spore-forming bacteria
(Ledenbach and Marshall, 2009).

CO₂ is Generally Recognized as Safe (FDA, 2011) and the dissolved gas is a natural
component of freshly drawn milk that is subsequently lost during processing and transit.
Hence, it can be utilized as a simple, inexpensive method to improve the shelf life of dairy
products. Since dissolved CO₂ is a natural component of freshly drawn milk that is
subsequently lost during processing and transit, there is an ample scope for its utilization as
a simple, inexpensive technique to extend shelf life of dairy products.

Daniels et al., (1985) postulated that the growth rate of aerobic bacteria was
hindered due to the displacement of oxygen by CO₂ in the product. Dixon et al., (1987)
proposed that interference by CO₂ in the biochemical pathways may adversely affect
microbial growth. In the beverage industry, carbonation leads to dissolved CO₂, which in
addition to its antimicrobial action, gives the product a sparkling appearance, astringency
and a refreshing aftertaste. In aqueous media, CO₂ forms carbonic acid that liberates H+,
resulting in a reduced pH and increased acidity of the beverage, creating a stress in the
microbial environment and adversely influencing its physiological activity (Wolfe, 1980;
Karagul-Yuceer et al., 2001).

Carbon dioxide is known to increase the acidity and meanwhile decrease the pH,
which could be due to production of carbonic acid upon carbonation (Roberts and Torrey,
1988). Pichard et al., (1984) reported that the role of carbon dioxide in inhibiting microbial
activity by alteration and regulation of enzymes. Carbon dioxide which is responsible for
taste tingling effervescence in beverages should be of high purity (Khurdiya, 1989).
Mitchell, (1978) reported that carbon dioxide provides the characteristic ‘sparkle’ to all
carbonated soft drinks and prolongs the shelf life of the product. Carbon dioxide has a
prevailing inhibitory effect on microbial growth (Dixon and Kell, 1989) and has been used
as a means for extending the shelf life of foods in technologies like controlled atmosphere
and modified atmosphere packaging (Kader, 1986). Carbonation is practiced in many
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 Review of Literature

beverages, like popular soft drinks, for the purpose of imparting a unique sparkle or
effervescence, flavor enhancement, and a refreshing aftertaste. The “fizz” generated in the
beverage due to dissolved CO₂ could position the dairy beverage as a novel and nutritive
beverage, appealing to the younger generation as a healthy alternative to carbonated soft
drinks. The antimicrobial action of dissolved CO₂ can be attributed to several factors.

Carbonation remarkably improved the sensory characteristics of the beverages, which

found to be more acceptable than the market sample (Meraj et al., 2009). As carbonation
improves colour and appearance, taste, consistency and overall acceptability, the
commercial beverages are invariably carbonated (Khurdiya, 1992). Carbonation of dairy
beverages is reported to improve its thirst quenching and refreshing quality, positioning
dairy beverages as a novel and nutritive beverage, appealing to the younger generation as a
healthy alternative to carbonated soft drinks. Carbonation has already been reported for
extension of shelf life of fluid milk / flavoured milk (Hotchkiss and Lee, 1996; Ravindra et
al., 2011) and dairy products like yoghurt (Karagul-Yuceer et al., 1999), ice cream mix
(Hotchkiss and Chen, 1996) and cheese (Mann, 1991).

Rajagopal et al., (2005), reported that the injection of carbon dioxide into the
milk beverage was observed to decreases the pH of the product, possibly due to the
dissolution of the gas and the resultant formation of carbonic acid. The carbonation of
drink had a significant effect on raising the acidity and lowering the pH of system without
adversely affecting the sensory quality of product. The carbonation inhibited the growth of
microbes, especially psychrotrophs, during storage, nearly doubling the shelf life (up to 30
days against 17 days for control) of beverage when stored under refrigeration. During its
refrigerated storage, the carbonated drink exhibited lower changes in FFA and soluble
nitrogen content than the control. The antimicrobial properties of CO₂ could be used as a
simple, inexpensive processing aid to extend shelf life of flavoured dairy beverage
(Ravindra et al., 2011). Singh et al., (2012) reported that carbon dioxide can reduce the
level of enzyme activity in raw milk; this minimizes the development of off-flavours
during storage and distribution.

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Choi and Kosikowski, (1985), also reported that the yeast and mold growth in
carbonated yoghurt beverage was suppressed due to carbonation. Carbonation did not
significantly alter the pH of the product while registering a marginal increase in the
titratable acidity of the product. Dissolved carbon dioxide aided in arresting the
development of lipolysis and proteolysis and inhibiting the growth of yeasts and molds in
the product during storage. The carbonated samples were found to be acceptable up to 12
weeks while the un-carbonated control sample kept well up to 5 weeks under refrigerated
storage. Dissolved CO₂ could be used as a simple and inexpensive processing aid to extend
shelf life of fermented dairy products (Ravindra et al., 2012).

2.5.6 STORAGE STABILITY AND SENSORY EVALUATION OF WHEY

BEVERAGE

Mandal et al., (1997) developed a formulation for beverage from fresh chhana
whey (deprotenized) by addition of 1.5 ml citric acid, 3 ml lemon juice and 10 per cent
sugar to 100 ml of diluted deprotenized whey. Shelf-life of the product was studied with or
without preservatives at room and refrigerated temperature for 45 days. A marginal
increase in viscosity and standard plate count was observed during the last stage of storage
period. Krishnaiah et al., (1989), was also reported that organoleptic quality of whey
beverages keeps reducing during the storage period.

The study was carried out to bioconversion of whey for preparation of beverage
was standardized by utilizing yoghurt cultures. The product, wheyghurt drink, made with
4per cent yoghurt cultures inoculated in deproteinized whey (4.8per cent lactose, 0.66 per
cent ash, 0.46 per cent fat and 0.40 per cent protein adjusted to pH 6.4) and incubated at
42⁰C for 8 hrs had all the technological requisite and dietetic criteria required in the
product. Incubation at 42⁰C with 4 per cent culture in whey exhibited highest inhibitory
activity. The product stored up to 5 days under refrigeration was of acceptable organoleptic
quality and requisite amount of microbial population (108 cfu/ml) to be potentially
beneficial (Kar and Misra, 1999).

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The carbonated whey beverage from chhana whey stored at refrigeration

temperature can be kept for longer duration (17 days) in contrast to the samples kept at
room temperature with respect to acceptable flavour, colour and appearance and overall
acceptability as well as pH (Pandey and Singh, 2002).

Suresha and Jayaprakasha, (2003), studied on development carbonated whey

permeate beverage. The beverage prepared with pineapple flavour was adjusted as the best,
followed by mango and orange. Carbonation remarkably improved the sensory
characteristics of the flavoured beverage. The non-carbonated beverages were found to
have shelf-life of about 15 days at 4⁰C. Whereas, carbonated beverage were found to have
shelf life of more than 30 days at similar storage conditions.

Saravanakumar and Manimegalai, (2005) studied on the storage stability of whey

based papaya juice blend ready to serve beverage. The beverage was prepared by blending
papaya juice at 10 per cent with whey. Total soluble solids (TSS) and acidity were
maintained at 15⁰Brix and 0.3 per cent respectively. During storage of 3 months at
refrigeration temperature there was an increase in acidity, reducing sugars and a decrease
in pH and TS did not change. A slight change in the microbial load was also observed.

Study was carried out on the development and storage of Whey Based Mango
beverage. The storability of the beverage was studied at 4±1ºC for 30 days (Ahmed et al.,
2011). The study revealed that the RTS beverage prepared with 70 per cent whey and 30
per cent mango juice scored maximum for almost all sensorial quality attributes such as
appearance, color, flavor, taste and overall acceptability and also found highest in ascorbic
acid content (9.80mg/100g) and the storage of beverage at refrigeration temperature over a
period of 30 days (Sakhale et al., 2012).

The study was conducted to develop a probiotic beverage using whey and
pineapple juice. Lactobacillus acidophilus was used as the probiotic organism. The level of
pineapple juice addition was optimized on the basis of sensory quality evaluation. The
study has revealed satisfactorily good quality probiotic beverage with therapeutic value
prepared by using a 65:35 blend of whey and pineapple juice inoculated with 1 per cent
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inoculum of Lactobacillus acidophilus with a shelf life of 24 days at 5±1°C and 48 hrs at
30±1°C (Shukla et al., 2013).

2.6 LASSI

2.6.1 DEFINITION
Lassi is one such a popular indigenous fermented milk beverage, originally from
Punjab, in India, made by blending yoghurt/dahi with water, salt and spices until frothy.
Lassi is a white to creamy-white, viscous liquid, with a sweetish, rich aroma and mild to
acidic taste. It is flavoured either with salt or sugar and other condiments depending on
regional preferences. Lassi is obtained from pasteurized milk or partly skim milk cultured
with lactic acid and aroma/ flavour producing organisms.
Originally, Lassi was the liquid left over from churning butter from milk, so it is butter
milk Lassi/butter milk widely known as “Chhachh” to be the liquid left over after
extracting butter from churned curd (dahi). Today, this is called traditional Lassi. On the
other hand artificially made, also known as cultured Lassi is a product, made by blending
yoghurt with water, salt, spices until frothy. Increasing awareness among consumers to
ensure good health coupled with the change in lifestyle has led to the concept of functional
foods. The development of functional foods is thus a unique opportunity to contribute to
the improvement of the quality of food and consumer health and well-being.
In the production of lassi, dahi is invariably diluted with water. The use of whey in place
of water as diluent would add to the nutritional superiority since it contains many health
promoting constituents. But use of unhydrolysed whey results in increased lactose content
in the lassi which may cause problem with lactose intolerant consumers. Hydrolysis of
lactose not only reduces the lactose content in whey but also increases sweetness of whey.

2.6.2 VARITATIONS

There are many types of lassi that are now available. Sweet lassi is a more recent
invention, and has become immensely popular. Rose water is a common ingredient for

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sweet lassi and adds a sweet, perfumed aroma. Sweet lassi can be flavoured with any fruit
of choice like mango, pineapple, banana, lychee, strawberry, etc. The traditional lassi is a
salty yoghurt drink which has a thicker consistency as compared to buttermilk. It can be
savoured with various spices and ingredients, but it almost always includes ground cumin
powder. Salty lassi is not only extremely easy and quick to make but also very refreshing
and cooling to beat the heat of summer.

a) Traditional mild sweet (or salty) lassi

This form of lassi is more common in the villages of Punjab and north India. It is
prepared by blending dahi with water and adding sugar and other spices to taste. Salt can
be substituted in place of sugar. The resulting beverage is known as salted lassi.

b) Sweet lassi

Sweet lassi is a form of lassi flavoured with sugar, rosewater and/or lemon, strawberry

or other fruit juice, saffron lassi, which are particularly rich, are a specialty of Sindh in
Pakistan, and Jodhpur and Rajasthan in India. Makkhaniya lassi is simply lassi with lumps
of butter in it (makkhan is the Gujarati, Hindu and Punjabi word for butter). It is usually
creamy like a milkshake.

c) Mango lassi
Mango lassi is gaining popularity worldwide. It is made from yogurt, water and mango
pulp. It may be made with or without additional sugar. It is widely available in UK,
Malaysia and Singapore, and in many other parts of the world. In various parts of Canada,
mango lassi is a cold drink consisting of sweetened kesar mango pulp mixed with yogurt,
cream, or ice-cream. It is served in a tall glass with a straw, often with ground pistachio
nuts sprinkled on top.

d) Bhang lassi
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Bhang (or bhung) lassi is a special lassi that contains bhang, a liquid derivative
of cannabis, which has effects similar to other eaten forms of cannabis. It is legal in many
parts of India and mainly sold during Holi, when pakoras containing bhang are also
sometimes eaten. Rajasthan is known to have licensed bhang shops, and in many places
one can buy bhang products and drink bhang lassi.

e) Banana Lassi
Banana lassi is prepared by mixing of crushed ice or ice water, natural yoghurt, very
ripe banana, peeled and roughly chopped and add honey (optional) with one pod crushed
cardamom and Blend all ingredients together in a blender. Served chilled and garnish with
a slice of banana.

f) Watermelon Lassi
Dorai et al., (2013) developed watermelon Lassi and the objective of this study was to
prepare traditional Lassi with the addition of a novel fruit component, water melon, in
powder form to make the product to have the natural flavour and sweetness from
Watermelon and the typical sourness from Dahi. The new product proved to be a perfect
thirst quencher with the natural flavour of watermelon. Sensory evaluation carried out on
the prepared product rated the new product as ‘highly preferred’ when compared to the
control plain lassi.

g) Whey-based Lassi

Whey based lassi, in which up to 60 % of milk is replaced with whey, has been
developed at National Dairy Research Institute, Karnal. The product formulation requires
addition of pectin, CMC and trisodium citrate. The product has 2.0 % fat, 1.8 %, 4.6 %
lactose and about 23.0 % total solids. This product can also be UHT-processed for long
self-life.

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Lassi like beverage was developed using paneer whey and buffalo milk with pectin
and CMC as stabilizers. After neutralization, paneer whey (pH 6.6) was mixed with
standardized buffalo milk (6% fat), inoculated with NCDC167 culture @ 1% and
incubated at 30°C for 14 to 16 h. Dahi was blended along with sugar syrup and flavour.
The beverage had 1.35% protein, 19 to 36% TS, 1.9% fat and 4% lactose. The product
with 70% whey and pectin, CMC in the ratio of 0.5:1.0, at 0.6 % level was adjudged most
acceptable (Mittal, 2003).

Lassi using buttermilk and soyabean was made by Deka et al. (1984). The blanched
soyabean cotyledons were ground with buttermilk to get slurry with soy-solids to
buttermilk solids ratio 2:1. The slurry (12%) was homogenized (175 and 35 kg/sq.cm at
65°C), pasteurized (85°C) and inoculated with L. bulgaricus and S. thermophiles @ 2%
after incubation at 37°C for 10 to12 h. Dahi was sweetened with sugar syrup and
thoroughly mixed. The resultant lassi had 10 days storage life at 5°C in polyethylene
pouches.

Attempt for enhancing the shelf-life of Lassi apart from cold storage was made by
Naresh et al. (1996). Addition of nisin @ 200 or 300 IU/ml and 400 or 500 IU/ml extended
the acceptability to 24 and 32 h respectively, compared to 8 h for control at 30°C.
Simultaneously at 20°C storage, the acceptability increased from 12 to 24 h and 48 h by
adding nisin @ 100 or 200 IU/ml and 400 or 500 IU/ml, respectively. Nisin @ 500 IU/ml
could extend acceptability up to 8 to 10 days at refrigerated storage.

2.6.3 Composition of lassi.

Lassi is scientifically described as a fermented milk beverage obtained after the

growth of selected culture, usually lactic streptococci, in heat treated milk followed by
sweetening with sugar. It is consumed as a cold, refreshing, therapeutic beverage usually in
summer months. It is prepared by breaking the curd into fine particles by agitation,
addition of sugar, water and flavour as desired. The composition of lassi is detailed below.
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Table 2.1 Composition of lassi (Aneja et al., 2003).

Milk fat 1.5-3.8 %

Milk TS 9%

Sugar 13-20 %

Sodium dihydrogen phosphate 0.5 %

Low methoxy pectin 0.5%

acidity 0.7 % LA (minimum)

2.6.4 HEALTH BENEFITS OF LASSI.

Lassi is a dairy product which is rich in potassium, calcium, protein and B-Vitamin
(B12). It is a great source of calcium for lactose intolerant people, as there is very little
lactose present in it and instead it contains the digestive enzyme, lactase.

The health giving and vitalising properties of fermented dairy products such as
Dahi, Yoghurt, Kefir and Koumiss have been documented since ancient times. In general,
fermentation plays a few specific roles in foods (Rashmi, 2010).

1) For successful application in foods, the probiotic used should also be

technologically compatible with the food manufacturing process.
2) In addition to these, foods containing the probiotic bacteria must maintain the
characteristic sensory attributes of the fortified traditional food.
3) It helps in maintaining the balance of gut flora which generally gets upset due to
metabolic disorders like constipation, diarrhea, dyspepsia, cystitis, chronic
ulcerative colitis and mucous colitis.
4) Intestinal gut flora exists in dynamic balance, but this may be disturbed in certain
conditions like prolonged diseases, food and water contamination and excessive use
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of antibiotics. Most of these health issues can be overcome by consuming probiotic

based dairy products.
5) Detoxification during food fermentation processing.
6) Enrichment of human diet through development of a wide diversity of flavour,
textures of food.
7) Bio-enrichment of foods substrates with protein, essential amino acids, essential
fatty acids and vitamins.
8) Lassi strengthens and stabilizes the immune system.
9) Its lowers LDL, raises HDL cholesterol.
10) Calcium rich lassi significantly increases body fat loss, boost the body’s ability to
build bone, helps to prevent and heal Arthritis.
11) Drinking lassi regularly can helps restore your good bacteria levels.
12) Preservation of foods via lactic acid, alcoholic and acetic acid fermentations.
13) Reduction in liberation of carcinogenic end products.
14) It helps in promoting digestive health and boosts the immune system.
15) It helps in preventing urinary tract infections.

Kumar (2004) observed that lassi is a popular fermented milk drink product that is
relished all over India. It is popular for its pleasant taste, cooling and thirst
quenching properties and therapeutic value. Keeping this in mind the product was
judged for its possibility of incorporating whey without compromising the sensory
attributes for people suffering from obesity and diabetes.

2.6.5 ADVANTAGES OF THE TECHNOLOGY 

 Provides an opening for the disposal of good quality returned surplus milk.
 Increase the profitability of dairy plants.
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 Requires minimum additional facilities.

 Amenable to large-scale production.

 Simple technology and does not require sophisticated equipments or machinery.

 Excellent quality milk based beverage

 Has high nutritive and therapeutic value

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