PROPOSAL ON

Development and Quality Evaluation of Whey-Based Banana Herbal Beverage

incorporating Ocimum sanctum (Tulsi)

PREPARED BY
Rachin Giri
B. Tech (Food) 4th year

Submitted To:
Department of Food Technology
Lalitpur Valley College
Institute of Science and Technology
Tribhuvan University, Nepal
Jawalakhel, Lalitpur
2023

Contents
1.Introduction.......................................................................................................................1
1.1.General Introduction ......................................................................................................1
1.2. Statement of the problem..............................................................................................3
2. Objectives …......................................................................................................................4
2.1 General objectives.........................................................................................................4
2.2. Specific objectives..........................................................................................................4
3. Significance of study..........................................................................................................4
4. Limitations of the study.....................................................................................................5
5. Materials and methods......................................................................................................5
5.1.Materials...........................................................................................................................4
5.1.1. Raw materials...............................................................................................................4
5.1.2.Apparatus......................................................................................................................5
5.1.3. Chemicalsrequired.......................................................................................................6
5.2Methods............................................................................................................................5
5.2.1 Preparation of Milk Whey.............................................................................................6
5.2.2 Preparation of banana juice …......................................................................................6
5.2.3 Preparation of ocimum sanctum extract…………………………………………………………………..7
5.2.4 Preparation of Whey Based Banana Herbal (WBBH) RTS Beverage…………………………..8
5.2.5Sensory analysis.............................................................................................................9
5.2.6 Physico- chemical analysis............................................................................................9
5.3 Analytical procedure…………………………………………..……………………………………………………….9
5.4 microbial alanalysis.........................................................................................................10
5.5 Data analysis…………………………………………………….………………………………………………………..10
5.6 Storage Stability………………………………………………………………..………………………………………..10
6. Expected Results…………………………………………………………………………………………….………………10
7. Estimation of expenses....................................................................................................11
8.Workplan…………………………………………..……………………………….……………………………………...11
9.Reference.........................................................................................................................11

Outline of the research work

TOPIC OF DISSERTATION: " Development and Quality Evaluation of Whey-Based

Banana Herbal Beverage incorporating Ocimum sanctum (Tulsi) "

Guide teacher: …………………………

Duration: Shrawan 2080 to Poush2070

Estimated expenditure: Rs. 21000

Laboratories: Laboratory of Lalitpur Valley College, Jawalakhel, Lalitpur.

Location: Jawalakhel, Lalitpur.

Objective: The main objective is to prepare of Whey-Based Banana Herbal Beverage

incorporating Ocimum sanctum (Tulsi)

Part I
1. Introduction
1.1 General Introduction:
The food industry is currently looking for ingredients which might provide good functional
and nutritional characteristics to manufacture different food items with value added. The
increased knowledge of consumers about nutrition, health, and food quality, as well as the
high level of competition in the market is forcing the food industry to search for these
ingredients. In this context, a group of novel functional food products has been developed
throughout recent years. Among these, over 40% of functional foods are dairy products and
fermented beverages containing milk whey are the principal functional beverages
(Ahmed,T.; Sabuz et. Al., 2023).
Whey is one of the highly nutritious by-products obtained from the dairy industry producing
cheese, chenna, and paneer. Use of paneer whey improved protein content of beverages
and showed highest mineral content among all the beverages suggesting use of paneer
whey based beverage as an electrolytic drink. Paneer whey not only improved the shelf life
and color of pineapple beverages, but also improved overall acceptability than cheese whey
(Baba, W. N., et al. 2016) Whey constitutes almost 45-50% of total milk solids; 70% of milk
sugar mainly lactose; 20% of milk proteins; 70-90% of milk minerals and most of the water-
soluble vitamins originally present in milk (Horton, 1995; Shukla et al., 2004). Whey contains
all the essential amino acids in excess to FAO standards e.g. isoleucine, lysine, threonine and
tryptophan. Whey consists of major proteins and minor proteins. Major whey proteins are
β-lactoglobulin-65%, α-lactalbumin-25%, serum albumin-8%, and minor proteins/peptides
are Glycomacropeptide(GMP), Bovine serum albumin, lactoferrin, Immunoglobulin,
Phospho-lipoproteins. Whey proteins have a Biological Value of 110, which is higher than
the value for casein, soy protein, beef, or wheat gluten, and have a high content of sulfur-
containing amino acids such as cysteine and methionine (Fox and McSweeney, 1998). Whey
proteins are high-quality proteins due to their high cysteine content and significant
quantities of B vitamins. They are also rich in calcium, phosphorous, magnesium, and zinc
(Hazen, 2005). Whey proteins are a rich source of branched-chain amino acids (BCAA) like
isoleucine, leucine, and valine. BCAAs unlike other essential amino acids are metabolized
directly into the muscle tissue and are the first amino acids used during periods of exercise
and resistance training (Shukla et. al., 2000).
After citrus, banana is the most important fruit in the world trade and is one of the most
widely grown fruit, cultivated over 170 countries along the tropic and sub-tropics of the
Capricorn. After ripening, banana fruit is highly susceptible to deterioration. A large quantity
of unmarketable surplus fruit is available in all the banana growing regions. Its considerable
amount is isted due to lack of efficient processing techniques that are unique to ripe
banana. Very few processed products are available in the market, primarily due to difficulty
in retaining the characteristics color, flavor and texture during the processing of ripe
banana.(Yadav et al. 2010, Dhamsaniya and Varshney 2013).

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Banana is good source of vitamin C, vitamin B6, potassium and dietary fibre. One banana
has 15% of the vitamin-C, 11% of potassium and 16% of dietary fibre for good health. It also
helps in protein metabolism, red blood cell formation and function of the central nervous
system. The vitamins C in banana also help to heal and defend against infection. Banana has
no fat, cholesterol or sodium. Bananas contain more digestible carbohydrates than any
other fruit. Banana is an important fruit which is used to cure many diseases. High in iron it
stimulates the production of hemoglobin in blood which helps in case of anemia. The
banana is used as a dietary food against intestinal disorder because of its soft texture and
smoothness. (Yadava, A., et al.2014)
Tulsi (Ocimum sanctum) is an aromatic plant which has many medicinal properties (Singh et
al., 2012). It contains several phyto-constituents such as cardinene, cubenol, borneol,
linoleic acid, orientin, linolenic acid, steric acid, oleic acid, palmitric acid, eugenol, vallinin,
vicenin, vitexin, vllinin acid, circineol, gallic Acid, vitamin A, vitamin C, phosphorus and iron
(Kadian and Parle, 2012) due to which it possess multifarious medicinal properties such as
antiviral, antifungal, antibacterial, antimalarial, anthelmintic, anti-oxidant, anti-cataract,
antiinflammatory, chemopreventive, radio protective, hepato-protective, neuro-protective,
cardioprotective, anti-diabetic, anti-hypercholesterolemia, anti-hypertensive, anti-
carcinogenic, analgesic, antipyretic, anti-allergic, immunomodulatory, anti-asthmatic,
diaphoretic, antithyroid, anti-fertility, anti-ulcer, anti-emetic, Anti-spasmodic, anti-arthritic,
adaptogenic, anti-stress, anti-cataract, anti-leukodermal and anti-coagulant (Cohen, 2014).
Fruits and vegetables tend to have very limited shelf life due to its perishable nature. In
order to preserve, they are processed to.These natural beverages are valued for their
nutritional content, refreshing quality, pleasant flavor and medicinal properties. Therefore,
blending of natural beverage is thought to be a good alternative for utilization and
preservation of fruits. Several types of beverages including blended, refreshing, and
functional beverages are prepared from fruits such as banana, apple, grapes, gooseberries,
litchi, pineapple, orange, etc. (Rathinasamy, M., et al. 2022).
Increased awareness in health issues leads to increase the consumption of fruit juices and
other natural products as an alternative to the traditional caffeine-containing beverages
such as tea, coffee or other soft drinks. Accompanying the increase in quantity of
consumption, there has been a parallel increase in the variety of fruit juices and beverages
offered for sale in the market. At present fruit beverages are generally synthetic flavored,
bottled and sold in the market. If this could be substituted with fruit juice and dairy whey, it
is more beneficial to the consumer, dairy industries and beverage manufacturers as well as
fruit growers(Dhamsaniya and Varshney 2013)). Therefore, it is very interesting to utilize the
milk whey as a water replacer for the development of a new delicious and nutritious RTS
beverage from the ripe banana juice. The use of milk whey produces off-flavor in the
beverages (Dhamsaniya and Varshney 2013). The Ocimum sanctum extract is used as
natural flavouring agent as its inclusion imparts a distinctive and appealing flavor profile,
enhancing the overall sensory experience of the beverage also it encompasses preservative
properties that contribute to the extended shelf life of the product. Beyond flavor and
preservation, tulsi is renowned for its medicinal properties, adding a potential health-

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promoting dimension to the beverage. Thus, Looking to the fast growing market segment of
functional beverages, it felt appropriate to use the ocimum sanctum extract tulsi in the
development of whey based beverage from ripe banana juice to fetch the higher market
demand.

1.2 Statement of the problem:

Whey is one of the highly nutritious by-products containing all 9 essential amino acids, is
obtained from the dairy industry producing cheese, chenna, and paneer. over the years,
several methods have been explored to convert huge volumes of whey into food-grade
products. The reason behind this is that only about 60% of whey generated globally is used
to manufacture products. The remainder of the material is disposed of as iste (León-López,
A., et al. 2020). Traditionally, whey has been discarded in surface water or fed to cattle.
Whey dumping on soil causes serious environmental pollutants by changing the physico-
chemical properties of soil, which leads to lower crop yields. Furthermore, whey iste can
also decrease dissolved oxygen and inhibit biodegradability when it is released into bodies
of water, and causes a serious threat to aquatic life and human health (T. Begum, et. al
2019). Considering the nutrient value and istage of milk whey, and environmental concern,
returning whey into the human food chain in a palatable way would be a more logical and
economical method than disposal.
Banana has a very short storage life. It is highly perishable, therefore susceptible to
several diseases resulting in extensive postharvest losses (Basel et al.,2002) and
considerable amount of this fruit is isted due to inadequate processing and preservation
techniques. Production of fruit juices, concentrates, and purees may be a good use for the
excess and rejected stocks (Maskan, M. 2000). It is utmost importance to process the large
amount of surplus ripe banana fruits available in the entire banana growing regions for
preventing their loss after ripening. Thus, Processing bananas into beverages reduces iste,
enhances flavor, and adds value. It creates convenient options, extends shelf life, and
promotes sustainability by utilizing surplus bananas effectively.
Ocimum sanctum, commonly known as Holy Basil, has a longstanding history in Nepal as a
therapeutic remedy. Its traditional use extends beyond therapeutic applications, as it
encompasses a rich repository of antioxidant and antimicrobial constituents that facilitate
the preservation beverages. Moreover, the herb offers an unmistakable aroma and flavor
profile, rendering it valuable as a flavoring agent.
By transcending its conventional role and undergoing purposeful processing, Ocimum
sanctum can undergo commercial integration into products like this. This strategic
transformation not only harnesses the herb's inherent potential but also engenders
diversification within the product portfolio. The infusion of this botanical constituent
augments the value proposition of the beverage, concurrently enhancing its status as a
functional consumable.
Through systematic processing and deliberate incorporation, the infusion of Ocimum
sanctum create new versions of the product and make it more valuable. his combination not

3
only uses the herb's natural properties but also helps make a unique drink that is good for
you. This shows a smart mix of old knowledge and what people want today.

2. Objectives:
2.1 General Objective
 To utilize Whey, ripe banana juice and Ocimum sanctum(Tulsi) for the preparation of
herbal beverage

2.2 Specific Objective

 To extract banana juice
 To analyze banana juice
 To extract Whey from milk
 To analyze Whey
 To prepare Ocimum sanctum extract
 To optimize the recipe of beverage
 To prepare Whey Based Banana Herbal(WBBH) beverage
 Sensory Analysis of WBBH beverage
 To analyze the physio-chemical characteristics of prepared final product.
 To analyze microbiological properties.
 To analyze effect of storage period on physiochemical properties and microbiological
properties.
 To carry out statistical analysis of obtained data.

3. Significance of study:
Investigating the formulation and quality of a functional beverage like Whey-Based Banana
Herbal (WBBH) beverage can contribute to promoting healthier dietary choices. This
beverage, enriched with the goodness of whey and Ocimum sanctum (Tulsi), has the
potential to offer unique health benefits, which could aid in disease prevention and overall
well-being.
Creating a novel beverage like WBBH beverage showcases innovation in the food and
beverage industry. By combining whey, ripe banana, and Tulsi, the study explores new
ways of utilizing natural ingredients to develop a unique and potentially nutritious product.
The utilization of whey, a byproduct of dairy processing, in the beverage industry is a
valuable step towards reducing iste and increasing its value. This study demonstrates an
alternative and beneficial use for whey, potentially encouraging further exploration of its
applications in functional foods and beverages.
The study adds to the body of scientific literature on functional beverages and food product
development. It provides empirical evidence of the potential benefits of incorporating whey
and Tulsi into a beverage, contributing to a deeper understanding of their interactions and
effects.

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4. Limitations:
 The different variety of banana is can be used to conduct the study.
 Storage effect on physiochemical and microbiological quality for longer days can be
study.
 Study on storage effect on physiochemical and microbiological quality on each
formulation can be done.

5. Materials and Methods:

5.1 Materials
5.1.1 Raw Materials
1) Banana Fruits:
 Fresh, ripe, banana is purchased from local market

2) Milk:
 DDC milk from local market

3) Ocimum sanctum (Tulsi) extract:

 Being a holy plant , is easily available in the market

5.1.2 Apparatus Required:

 Electronic Balance
 Measuring Cylinder, beaker, pipette, Volumetric flask,Test tube, conical flask,
burette
 Muslin cloth
 PH meter
 Refractometer Chemical Required:
 Thermometer  Citric acid
 Knives  NaOH
 Grinder  Sulphuric acid(H2So4)
 Stainless steel cooking pot  Amyl alcohol
 Crown corking machine  Fehling's solution
 Glass bottles (200ml)  plate count agar
 potato dextrose agar
5.2 Methods:
5.2.1 Preparation of milk whey:
The milk is boiled in a stainless steel vessel at 80°C for 10 minutes. The boiled milk after
cooling to 71°C is acidified by adding 2 (2g per kg of milk) citric acid (Eagle brand) followed
by continuous stirring until the coagulation of milk proteins (casein) taken place. The
mixture of milk whey and casein is allowed to cool at room temperature and then the whey

5
is filtered using the muslin cloth. The milk whey thus obtained is filled in a glass bottle and
stored in the refrigerator (7 ± 1°C) for further use.

0.2 % of citric acid(2g per kg)

5.2.2 Preparation of banana juice:

Natural banana juice is prepared without imparting any chemical/ enzymatic treatments as
per the method suggested by Dhamsaniya (2012). The process flow diagram for the
preparation of natural banana juice is shown in Figure 1. Ripe bananas are ished with
chlorinated water followed by manual peeling and cutting into slices of 3 – 4 mm. The
banana slices along with water in the ratio of 1:2 (banana slices: water, w/w) are heated at
100℃ for 45 minutes followed by cooling, pressing, filtration, pasteurization (at 90℃ for 10
min) and again cooling at room temperature. The prepared banana juice is stored at 7±1℃
in the refrigerator for further use.

6
 is

is

5.2.3
Extraction of Basil:
Juice The holy basil are properly and blended in a laboratory blender to paste with distilled
water (1:8) and filtered through muslin cloth to obtain the juice extract(Chauhan et al.
2012).

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5.2.4 Preparation of Whey Based Banana Herbal (WBBH) RTS Beverage:
Whey Based Banana Herbal(WBBH) beverages are prepared with the addition of Ocimum
sanctum extract following the method followed by Sirohi et al. (2005). For preparation of
beverage, Whey amount varying from 72 to 81 ml is added with fruit juice of varying amount
from 10-15 ml and 1-3% of ocimum extract as given in Table :. They whey, banana juice and
sugar are mixed in the given amount, preheated to 45oC before mixing ocimum extract as
shown in Fig:1. The beverages obtained are filtered and filled hot into pasteurized glass bottles
of 200ml. The filled bottles are pasteurized at 72C for 30 min and let cool down to room
temperature (28 C). Sensory analysis is done for all prepared compositions, and based on
sensory score best sample is selected for further chemical and microbiological analysis (Singh,
2014).
Table: Recipe for the preparation of 100 ml of WBBH beverage.

Treatment Banana juice (ml) O. sanctum Whey (ml) Sugar (g)

extract (ml)
A 10 1 81 8
B 10 3 79 8
C 10 5 77 8
D 15 1 76 8
E 15 3 74 8
F 15 5 72 8
Whey

Filtration (to remove foreign material)

Banana juice Heating at 45oC Sugar

Ocimum sanctum extract Mixing

Filtrarion Through Muslin Cloth

Bottling and crown corking (200 ml glass bottle)

Pasteurize (72oC for 30 min, boiling water) & cool at room temperature

Sensory evaluation

Selection of best formulation

Physio-chemical analysis of beverage (pH, acidity,TSS)

Figure 1: Flow chart for the preparation of a whey-based

banana beverage

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5.2.5 Sensory analysis:
The prepared beverage samples with varying levels of banana juice, O. sanctum extract and
milk whey are presented to a panel of ten semi-trained at comprising of faculty members and
undergraduate students. The panelists are associated with the research and development of
new food products at the department of food technology , Lalitpur Valley College, jawalakhel,
Lalitpur. A nine-point Hedonic scale score card is provided to the panelists to adjudge the
quality of the product with respect to colour, appearance, taste, flavor and overall
acceptability. The best-scored sample is s then selected for further chemical and
microbiological analysis and stored for 30 days in a refrigerator (7±1°C) for shelf-life study.

5.2.6 Physio-chemical analysis:

The physicochemical properties such as total soluble solids, fat content, acidity, pH, ascorbic
acid, and microbiological analysis are done for raw whey, banana juice, and prepared
beverage. The proximate compositions such as protein content, crude fiber, total and reducing
sugar, ash content, are determined (Ranganna, 1986).
The above mentioned analysis are performed for 20 days in 5 days interval to check the
shorage effect in refrigeration temperature and ambient room temperature final product is

5.3 Analytical procedure

i. Tritable acidity
Titration is carried out by titrating clear juice with standard 0.1N NaOH (AOAC 1995) and result
is expressed in terms of lactic acid for whey and citric acid for banana juice and final product.
ii. Proximate analysis
The proximate analysis of energy drink is carried out(Ranganna, 1986) in laboratory. iii. Protein
content
iv. PH
It is directly measured by using ph meter which is standardized by using buffer
solution of ph 7 and 4 at temperature required.

v. Total Soluble Solid (TSS)

TSS is determine by using refractometer and the values are expressed as oBrix.
vi. Moisture content
Moisture content of sample is determined by evaporating the water from the juice by
placing in a petri plate in an incubator(Ranganna, 1986)

9
vii. Total solids
Total solids of juice sample is determined by subtracting the moisture from the 100
(Ranganna, 1986).
viii. Fat%
Fat content is determind by Gerber centrifuge method (NDDB, 2001)

5.4 Microbial analysis

Total plate count (TPC) is determined by pour plate technique on plate count agar (PCA)
medium (incubated at 30o/48h). Yeast and molds count is determined by pour plate technique
on potato dextrose agar (PDA) medium (incubated at 25-27oC/48-72h).
The microbial test is also carried out in 5 days interval for 20 days to see the storage effect on
product's microbial quality while storing in ambient room temperature and refrigeration
temperature.

5.5 Data analysis

Different kind of data visualization is done for obtained data and Analysis of variance (ANOVA)
is carried out for data from sensory evaluation.

5.6 Storage stability

The prepared beverage is filled in glass bottles in aseptic condition. For the shelf-life
determination product is placed under ambient and refrigerated condition. Different
physiochemical parameters are observed under 20days interval.

7.Expected results
Whey based banana herbal beverage is prepared which is useful for industries to prepare new
variety of beverage utilizing the byproducts like whey and local valuable herb like ocimun
sanctum. Also it is expected that the beverage prepared will have medicinal value to some
extent with distinctive taste and flavour. Moreover, this work also study the shelf-life
extension of whey based fruit beverage by addition of herbal extract..

10
8. Estimation of expenses
1. Transportation and communication 2000
2. Stationary 2000
3. Raw materials and Chemicals 10000
4. Report writing and blending 5000
5. Research station overhead 500
6. proposal presentation 100
7. Miscellaneous 1000
Total 21000

9. Work Plan
The research work is supposed to be completed within 4 months from the date of
approval.Time line with research activities is as mentioned

Activities/Months Bhadra Ashoj Kartik Mangsir

1. Adjustment of
requirements
2. Preparation of the
product
3. Analysis
4. Data analysis
5. Report writing
6. Submission of final
report

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Professional. londo
Hazen, C. (2005). Packing that Protein Punch. Food Product Design.15:4, 32-55

Shukla, F. C., Sharma, A. and Bakshi, A. K. (2000). Studies on the preparation of fruit juice beverages
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Basel, R. M., Racicot, K., & Senecal, A. G. (2002). Long shelf life banana storage using MAP storage
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