PROJECT REPORT

FORMATION SYMBIOTIC ICECREAM

FROM COTTON SEED MILK
Submitted by

KISHORE SRINIVASAN R(202FD124)

ASWINTH K (202FD108)
SUBIKSHON(202FD150)

In partial fulfilment for the award of the degree

of

BACHELOR OF TECHNOLOGY
In
FOOD TECHNOLOGY

BANNARI AMMAN INSTITUTE OF TECHNOLOGY

(An Autonomous Institution Affiliated to Anna University, Chennai)
9ATHYAMANGALAM-638401

1
 ANNA UNIVERSITY: CHENNAI 600 025

OCTOBER 2023
PROJECT REPORT

FORMULATION OF SYMBIOTIC
ICECREAM FROM COTTON SEED MILK
Submitted by

KISHORE SRINIVASAN R (202FD124)

ASWINTH K (202FD108)
SUBIKSHON(202FD150)

In partial fulfilment for the award of the degree

of

BACHELOR OF TECHNOLOGY
In
FOOD TECHNOLOGY

9` 89ol,.q .

BANNARI AMMAN INSTITUTE OF TECHNOLOGY

(An Autonomous Institution Affiliated to Anna University, Chennai)
SATHYAMANGALAM-638401
Z3SSSa

2
 ]UYANNA UNIVERSITY: CHENNAI 600 025
OCTOBER 2023

. BONAFIDE CERTIFICATE

Certified that this project report FORMULATION OF SYMBIOTIC ICE CREAM

FROM COTTONSEED MILK is the Bonafide work of “KISHORE
SRINIVASAN R(202FD124), ASWINTH K(202FD108) AND
SUBIKSHON(202FD150)” who carried out the project work under my
supervision.

SIGNATURE SIGNATURE

Mr. GOWRISHAKAR PIDATHALA SAHITHYA

HEAD OF THE DEPARTMENT ASSISTANT PROFESSOR – II
Department of Food Technology Department of Food Technology
Bannari Amman Institute of Technology Bannari Amman Institute of Technology

Submitted for Project Viva Voice examination held on ………………….

Internal examiner 1 Internal examiner 2

3
 DECLARATION

We affirm that the project work titled FORMULATION OF SYMBIOTIC ICE

CREAM FROM COTTONSEED MILK being submitted in partial fulfillment for

the award of the degree of Bachelor of Technology in Food Technology is the

record of original work done by us under the guidance of MS. PIDATHALA

SAHITHYA, Assistant Professor - II, Department of Food Technology. It has
not formed a part of any other project work(s) submitted for the award of any
degree or diploma, either in this or any other University.

KISHORE SUBIKSHON B ASWINTH K

SRINIVASAN R (202FD150) (202FD108)
(202FD124)

I certify that the declaration made above by the candidates is true.

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 Ms. PIDATHALA SAHITHYA

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 ACKNOWLEDGEMENT

We would like to enunciate heartfelt thanks to our esteemed Chairman

Dr. S. V. Balasubramaniam, and the respected Director Dr. M. P.
Vijayakumar, for providing excellent facilities and support during the course
of study in this institute.
We are grateful to GOWRISHANKA R, Head of the Department,
Department of Food Technology for his / her valuable suggestions to carry out
the project work successfully.
We wish to express our sincere thanks to Ms. PIDATHALA SAHITHYA,
Assistant Professor - II, Department of Food Technology for his constructive
ideas, inspiration, encouragement, excellent guidance, and much-needed
technical support extended to complete our project work.

We would like to thank our friends, faculty, and non-teaching staff who
have directly and indirectly contributed to the success of this project.

ASWINTH (202FD108)
KISHORE SRINIVASAN R (202FD124)
SUBIKSHON B (202FD150)

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 ABSTRACT

Cotton seed milk, also known as paruthi paal in Tamil Nadu, India, is a nutritious drink. It's a
"riple nutrient" because it's high in protein, essential fatty acids, and sugar. It is also
considered an energy drink. Cotton seed milk is underappreciated despite its high protein
content. We used cotton seed milk (plant-based) to make ice cream because it is a good
alternative to animal-based milk. We're taking the initiative to reach out to city dwellers by
turning it into ice cream. Cotton seed milk has 23.1g of protein, 22.9g of fat, 43.2g of
carbohydrates, and 7.3g of fiber, making it an excellent choice for ice cream. It improves
brain function in children, aids in menstrual cycle regulation, produces enough insulin, heals
stomach ulcers, and increases breast milk production. For the preparation of ice cream,
cottonseed milk, and rice flour are combined to create a smooth and creamy texture for the
ice cream. The maple syrup improves the flavor and sweetness of the ice cream. Ice cream's
physiochemical and sensory properties are assessed.

KEYWORDS: Anti-oxidant, Capsaicin, Chilli powder, Pu

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 TABLE OF CONTENTS

Ch. No Title Page no.

1 Introduction 11
1.1 A drawback of milk-based ice cream 13
1.1.2 Environmental impact 13
1.2 Uses of Probiotic
1.3 Extraction of milk from cottonseed 13
1.3.1 Analysing of physiochemical properties of cottonseed milk
2 Literature survey 15
3 Objectives and methodology
3.1. Objectives 20
3.1.1. To study the toxicity of cottonseed
3.1.2 Removing gossypol toxicity from cottonseed
3.1.3 formulation of ice cream from cottonseed milk
3.1.4 Adding Symbiotics into the cottonseed ice cream
3.2. Methodology 22
3.2.1 Extraction of cottonseed milk 22
3.2.2 Chemical analysis of cottonseed milk 24
3.2.2.1 Examination of fat in cottonseed milk 25
3.2.2.2 Examination of cottonseed milk's protein content 27
3.2.2.3 Analyzing the moisture content of cottonseed milk 27
3.2.2.5 Examination of water activity in cottonseed milk 27
3.2.2.6 Quality parameters and microbial analysis
3.2.2.7 pH METER FOR TESTING pH
3.3.2.8 Acidity test 28
5.2.2.11 color analysis of cottonseed milk 28
3.2.2.9.1 Incorporate Symbiotic Elements: 29
3.2.2.9.1 Cool and Freeze: 30
3.2.2.10 Incorporating symbiotic components into ice cream 31
4 Proposed work modules 32
4.1. Ingredient 32
4.2. Formulation of ingredients 32
4.3. Blending of Ingredients 33
4.4. Processing of Vegan egg 33
4.5. Nutritional analysis of Vegan Egg 34
4.6. Packaging 34
4.7. Shelf-life studies 35
5 Results and discussions 36
5.1 Extraction of cottonseed milk 36

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 5.2 Chemical analysis of cottonseed milk 37
5.2.1 Fat Analysis Using Gerber Method
5.2.2 Protein analysis using the Kjeldahl technique
5.2.3moisturecontent
5.2.4 PH analysis
5.2.5 Color analysis of cottonseed milk
5.3 chemical properties of symbiotic ice cream from cottonseed
milk
5.4 Comparison of cow milk and cottonseed milk
5.4.1 Comparison graph of cow milk and cottonseed milk
5.4 cost analysis of symbiotic ice cream from cottonseed milk

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 LIST OF FIGURES

Fig. 1.1 A drawback of milk-based ice cream 11

Fig. 1.2 Kashmiri chilli, Wrinkled chilli, Chappatta chilli 12
Fig. 1.3 Process flow of hard-boiled vegan egg 13
Fig. 3.2.1.1 Alkali extraction of SPI 23
Fig. 3.2.1.2 Process flow of alkali extraction of SPI 23
Fig. 3.2.2.1 Formulation and optimization of vegan egg white 24
substitute
Fig.3.2.2.2 Formulation of vegan egg white 25
Fig. 3.2.3.1 Formulation of vegan egg yolk 26
Fig. 3.2.3.2 Formulation and optimization of vegan egg yolk 26
substitute
Fig. 3.2.4.1 Incorporation of vegan egg yolk substitute with egg 27
white
Fig. 3.3.1.1 Protein analysis using kjelTRON 29
Fig. 3.3.2.1 Fat analysis 30
Fig. 3.3.3.1 Fibre analysis 31
Fig. 3.3.4.1 Humidity chamber 31
Fig. 5.1.1 Textural analysis of vegan egg yolk 36
Fig. 5.1.2 Textural analysis of vegan egg white 37
Fig. 5.2.1 Nutritional content of chicken egg 37
Fig. 5.2.2 Nutritional content of vegan egg 38
Fig. 6.1 Hard-boiled vegan egg 40

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 CHAPTER – 1

INTRODUCTION
Cotton seed milk, manufactured from the seeds of the cotton plant, is a dairy-free substitute
for regular cow's milk. It is a great source of protein, calcium, and other vital elements, with a
creamy and nutty flavor. For people who are lactose intolerant, vegan, or just trying to cut
back on their intake of animal products, cotton seed milk is a fantastic substitute. It may be
substituted for cow's milk in several recipes, such as those for smoothies, porridge, and baked
goods.
Cotton seed milk is prepared by soaking the seeds, blending them with water, then straining
them to get rid of any solids. The resultant milk has a little nutty flavor that goes well with
both savory and sweet foods. It is smooth and creamy. All things considered, cotton seed milk
is a delightful and nourishing substitute for conventional dairy milk that will delight your
palate and provide you with all the vital nutrients your body needs. Try it now to experience
the silky sweetness of cottonseed milk.

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Fig. 1.1Cottonseed
Packed with nutrients: Protein, vitamin E, and minerals like calcium and magnesium may all
be found in excellent amounts in cotton seed milk. These nutrients are necessary for good
health and well-being in general. Lactose-free: For those with dairy allergies or lactose
intolerance, cotton seed milk is an excellent substitute. For individuals who are unable to eat
conventional cow's milk, it offers a dairy-free alternative. Heart-healthy: Cotton seed milk is
a heart-healthy option since it is low in cholesterol and saturated fat. It can lessen the risk of
heart disease and help lower cholesterol levels. Encourages weight management: Cottonseed
milk is a wonderful choice for people trying to control their weight since it has less calories
than cow's milk. Additionally, it has a lot of protein, which helps maintain a feeling of
fullness and satisfaction.

Environmentally friendly: Compared to conventional dairy milk, cotton seed milk is a more
environmentally friendly and sustainable alternative because it is produced from a byproduct
of cotton cultivation. Adaptable: You may use cottonseed milk in a wide range of recipes,
such as savory foods, baked items, and smoothies. Its mild taste goes well with both savory
and sweet foods. Tamil Nadu is now able to purchase cottonseed milk, a tasty and nutritious
substitute for regular dairy milk. Cottonseed milk, which is made from the squeezed seeds of
cotton plants, is a fantastic source of vitamins, minerals, and protein. Moreover, it has no
lactose, so anyone with dairy allergy or lactose intolerance can use it. There are several
applications for cottonseed milk, such as smoothies, coffee, cereal, baking, and cooking. It
goes well with both savory and sweet foods because of its somewhat nutty taste and creamy
texture. Try cottonseed milk if you're seeking for a locally produced dairy-free milk
alternative in Tamil Nadu. It is an eco-friendly and sustainable decision that encourages the
use of substitute plant-based goods and helps regional farmers. In Tamil Nadu, look for
cottonseed milk in specialist food stores or local grocery stores.

Live bacteria known as probiotics, when taken in sufficient quantities, offer many health
advantages. They support digestive health and a healthy balance of gut bacteria, which is why
they are frequently called "good" or "friendly" bacteria. Probiotics are present in a number of
foods, including kefir, sauerkraut, yogurt, and nutritional supplements. Probiotics have been
researched for their ability to strengthen immunity, facilitate better digestion, and promote
general health. Probiotics may be able to lessen the symptoms of digestive diseases such
diarrhea, inflammatory bowel disease (IBD), and irritable bowel syndrome (IBS), according
to research. Allergies, yeast infections, and urinary tract infections may also be prevented and
treated with their assistance. Probiotics help gastrointestinal health, but they've also been
connected to better mental and cognitive wellness. The gut-brain axis, which links the two
organs, may be involved in mood and behavior regulation, according to some research.
Probiotics may help reduce the symptoms of anxiety, depression, and other mental health
issues by supporting a balanced population of gut bacteria.

Prebiotics improve gut health by stimulating the development of good bacteria, strengthening

12
the immune system, and helping with weight control through effects on satiety and
metabolism. They are essential to the immune system's function. Enhancing mental health:
Prebiotics may enhance mental health by fostering a healthy gut microbiota. The gut-brain
axis is a complex communication link between the stomach and brain. Supporting nutrition
absorption: By encouraging the development of helpful bacteria that aid in digestion and
nutrient absorption, prebiotics can help enhance nutrient absorption in the gut. Managing
gastrointestinal diseases: By encouraging the development of beneficial bacteria and
enhancing gut health, prebiotics may be helpful for people with specific gastrointestinal
illnesses, such as irritable bowel syndrome (IBS) or inflammatory bowel disease (IBD).
Encouraging general health and well-being: Adding prebiotics can improve general health
and well-being by strengthening immune system function, improving gut health, and
facilitating nutrient absorption.

1.1 A drawback of milk-based ice cream

While many people prefer milk-based ice cream, there are a few drawbacks to it. Lactose
intolerance: Lactose, a sugar present in milk, is one of the main drawbacks of milk-based ice
cream. After drinking milk-based ice cream, those who are lactose intolerant may feel
digestive discomfort such as gas, bloating, and diarrhea. Allergies: One of the most common
allergens is milk, and for those who are sensitive to milk, ice cream containing milk might
exacerbate their symptoms. In certain instances, the symptoms might vary from moderate
hives or itching to severe anaphylaxis. Saturated fat and cholesterol: When taken in excess,
milk-based ice cream's high levels of saturated fat and cholesterol can lead to cardiovascular
problems. Frequent eating of high-fat foods, such as ice cream, can raise the risk of heart
disease and cause weight gain. Calories and sugar: Regular ice cream made from milk is
frequently heavy in both of these nutrients, which can be troublesome for people trying to
control their weight or who have diabetes. Tooth decay and other health problems can also
result from consuming sugary foods in excess.

1.1.2 Environmental impact:

Dairy production, including milk for ice cream, can have significant environmental
consequences such as greenhouse gas emissions, water pollution, and deforestation for
grazing land. This environmental impact contributes to concerns about sustainability and
climate change. Animal welfare: The production of milk for ice cream involves the dairy
industry, which has raised concerns about animal welfare practices such as confinement,
separation of calves from their mothers, and routine use of antibiotics and hormones.
These drawbacks emphasize the need for moderation and the exploration of substitutes, such
as plant-based ice creams that may allay some of these worries and are prepared with
components like almond, coconut, or soy milk.

1.2 Uses of Probiotic

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One kind of probiotic bacteria that is frequently seen in the human intestine is Lactobacillus
acidophilus. It is frequently used as a dietary supplement to support gut health since it is
essential for preserving a balanced population of bacteria in the digestive tract. A summary of
Lactobacillus acidophilus will be given in this introduction, along with information on its
origins, advantages, and possible drawbacks. Additionally, we'll talk about how it might be
utilized to promote general health and well-being.
1.3 Extraction of milk from cottonseed
The procedure of extracting milk from cotton seeds entails isolating the component that
resembles milk from the other ingredients. This nutrient-rich, milk-like material can be
utilized as a nutritional supplement or in the manufacturing of cotton seed oil, among other
applications. To release the milk-like fluid, the cotton seeds are usually crushed during the
extraction process. After that, this material is extracted from the solid components using
centrifugation or filtration techniques.
Cotton seed milk is a rich resource that finds use in the food and beverage, cosmetics, and
pharmaceutical sectors. To increase milk supply and preserve milk quality, it's critical to
make sure the extraction procedure is carried out safely and effectively.

1.3.1 Analysing of physiochemical properties of cottonseed milk

Consumers seeking dairy-free choices are increasingly turning to cottonseed milk, a plant-
based substitute for conventional dairy milk. Made from cotton plant seeds, this milk is
frequently enhanced with vitamins and minerals to replicate the nutritional makeup of cow's
milk. Analyzing the physicochemical characteristics of cottonseed milk is crucial to
comprehending its attributes. The physical and chemical qualities of a material, such as its
pH, viscosity, density, and protein content, are referred to as its "physicochemical
properties." A significant physicochemical characteristic of milk made from cottonseed is its
pH level. A substance's pH value indicates how acidic or alkaline it is; a pH of 7 indicates
neutrality. Depending on the processing techniques used to manufacture it, cottonseed milk
can have a pH of 6.5 to 6.8, which is normally mildly acidic.
The viscosity, or thickness, of cottonseed milk is another crucial characteristic. The
temperature of the milk as well as its protein and fat content can affect its viscosity.
Compared to cow's milk, cottonseed milk often has a lesser viscosity, which might have an
impact on the mouthfeel and texture.
Another crucial physicochemical factor to take into account is the milk's density from
cottonseed. Density, which is defined as a substance's mass per unit volume, affects how milk
behaves and interacts with other components in recipes. Generally speaking, cottonseed milk
is less dense than cow's milk, which might have an impact on how well it foams or emulsifies
in baking and culinary applications. Another important physicochemical characteristic of
cottonseed milk is its protein content. An essential nutrient, protein is involved in several
bodily physiological functions. Protein is frequently added to cottonseed milk to increase its
nutritional content and make it a more complete dairy substitute. To sum up, it is critical to
examine the physicochemical aspects of cottonseed milk to comprehend its attributes and its
applications. Through the examination of characteristics like pH, viscosity, density, and
protein content, scientists and producers may enhance the caliber and performance of

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cottonseed milk as a substitute for dairy products.

CHAPTER - 2
LITERATURE SURVEY

Feeding soy products with oil reduced the amount of casein and milk protein; this was not the
case with cottonseed. When oil or entire seeds were added, the amount of feed consumed
decreased. Free oil decreased the overall tract's capacity for digestion. As a result of
providing free oil, the proportion of propionate and butyrate in ruminal VFA was greater and
lower, respectively. Protozoal counts decreased when whole seed and free oil were added, but
they remained unchanged when roasted oil seed was added. Rumen fluid's unsaturated long-
chain fatty acid concentration rose in response to free oil supplementation. Milk protein
percentage and the ratio of blood glucose to unsaturated long-chain fatty acids were
positively associated.Grummer et al. (2008), RR

Cows in early first lactation and older cows in either early or late lactation were assigned to
separate squares in the Latin square based on their production status. Ad libitum diets were
provided, with 21-day intervals between meals. With cottonseed feeding, the percentages of
milk fat and total solids rose but the amount of protein fell. When fed cottonseed, milk casein
nitrogen dropped from.387 to.375%. Whey's nitrogen content rose while non-protein nitrogen
stayed constant. The amount of nonprotein nitrogen in the casein and whey fractions rose,
while the amount of total nitrogen remained same.DePeters, EJ, et al., 1995

Milk from cottonseed was extracted by soaking 100 g of cottonseed in 600 ml of water for 90
minutes at 32 C. In a blender, soaked cottonseeds were mixed for 90 seconds. Samples of
cottonseed slurry were treated individually with 1% protease, α-amylase, and cellulase
enzyme at pH 7.0. The samples were incubated at 40 and 52 degrees Celsius, and the enzyme
treatment took 2.30 hours to fully hydrolyze the slurry samples. The cottonseed milk was
packaged in a 250 ml flask and kept at 4 C for additional analysis after the slurry was filtered

15
through muslin cloth and the enzymes were inactivated at 90 C for five minutes.(S.
Vellaikumar & others, 2021).

Thirty-two primiparous and 12 multiparous Holstein cows were randomly assigned at calving
to treatments to determine the effects of type and amount of cottonseed product on plasma
gossypol, milk production, and composition, and conjugated linoleic acid concentration in
milk fat. Rations consisted of corn silage, corn grain, soybean meal, and cottonseed hulls, and
contained on average 16.8% crude protein and 25.3% acid detergent fiber on a dry matter
basis.(SM Noftsger et al., 2000)

From June to August 2011, the current investigation was carried out at the Livestock
Research and Development Station Surezai, Peshawar. The purpose of this study was to
ascertain how Cotton Seed Cake (CSC) affected the live weight, body condition score (BCS),
feed intake, milk output, and milk composition of Holstein Friesian (HF) crossbred cows
(Khrshaid Anwar et al., 2013).

Dairy products like ice cream have a strong chance of serving as a food source for probiotic
microorganisms. The general public consumes ice cream, which in addition to being a
functional healthy meal, contains helpful ingredients including dairy products, vitamins, and
minerals. For these reasons, adding probiotic bacteria to ice cream is extremely useful.
Additionally, ice cream significantly increases the vitality of probiotic strains throughout
production—especially during storage—when compared to fermented milks as a carrier.
(2011) Reza Mohammadi and colleagues

three types of ice cream: prebiotic (made with inulin, a prebiotic substrate); synbiotic (made
with probiotic microorganisms and inulin combined); and probiotic (made with potentially
probiotic microorganisms like Lactobacillus casei and Lactobacillus rhamnosus). The ice
creams' pH, acidity, and functional and physical qualities were assessed in addition to their
microbiological counts. After being frozen, the probiotic bacteria in the experimental ice
creams were maintained, and their viable lactic acid bacteria counts reached the minimal
level needed to provide probiotic effects. A Fratianni and associates, 2010

This study used calcium alginate and whey protein concentrate microencapsulation to assess
the survival of three probiotic strains in ice cream: Lactobacillus plantarum, Lactobacillus
casei, and Bifidobaterium bifidum. Additionally, the effects of lactulose,
fructooligosaccharides, and inulin were investigated. 2% fructo-oligosaccharides were used
in the production of six different forms of synbiotic ice cream that contained free and
microencapsulated L. plantarum, L. casei, and B. bifidum. Even in the presence of FOS, the
survival of all tested strains increased with varying sugars. Additionally, compared to loose
cells, the vitality of microencapsulated strains improved by more than one log cycle.El-
Sayed, Hoda S. et al. (2014)

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A study was carried out to prepare ice cream by incorporating Lactobacillus acidophilus and
Saccharomyces boulardii as probiotics and fructo-oligosaccharides (FOS) as a prebiotic and
enriched with whey protein concentrate and analyzed for the viability of probiotics during
storage of ice cream and the effect FOS on survival of probiotics in human gut. The protein
content and melting quality of the ice cream samples showed a significant (P< 0.01)
difference. The FOS in the ice cream mix significantly (P< 0.01) improved the growth of
probiotic cultures. A significant reduction in the count of L. acidophilus and S. boulardii was
observed during storage of ice cream. Consumption of synbiotic ice cream significantly
increased faecal L. acidophilus and S. boulardii counts in human volunteers. The L.
acidophilus and S. boulardii count were higher in the treatments when L. acidophilus and S.
boulardii were incorporated in combination than alone(TR Gopalakrishnamurthy et al., 2012)

Prebiotics are substances that can promote the growth of beneficial microorganisms, mainly
in the intestinal tract, and will modify the colonic microbiota. The following health benefits
are attributed to prebiotics: relief from poor digestion of lactose, increased resistance to
bacterial infection, better immune response and possible protection against cancer, reduction
of the risk of diseases such as intestinal disease, cardiovascular disease, non-insulin
dependent diabetes, obesity and osteoporosis. This article presents a discussion of prebiotics,
with descriptions of the concepts and its use in clinical practice, and a review of some recent
research showing the benefits that these ingredients provide to human health and providing
data on the recommended intakes for consumption.( Sgarbieri Valdemiro Carlos et al., 2019)

Probiotics and prebiotics play an important role in human nutrition. In recent years there has
been a significant increase in research on the characterization and verification potential health
benefits associated with the use of probiotic and prebiotic. The main effects attributed to
selected probiotics/prebiotic products have been proved by clinical trials, while others have
been acquired on the basis of in vitro tests which require in vivo transposition in order to be
validated. The main clinical reports in the literature for the application of probiotic have been
done for the treatment of infectious diseases including viral, bacterial or antibiotic associated
diarrhoea, relief of chronic bowel inflammatory diseases, immuno-modulation, lowering of
serum cholesterol, decreased risk of colon cancer, improve lactose digestion, reduce allergies,
and effect on intestinal microbiota.(PHILIPEE Bressollier et al., 2020)

In today's well-resistant pathogens and excessive use of antibiotics which weake and
undermine the immune system the importance of pre-and probiotics is more desired.
Probiotics-lactic acid bacteria-our intestinal symbiotes, has significant affect on our intestinal
tract and brings us to number of positive physiological effects-inhibit the development of
pathogenic microflora and serious stimulate the immune system, which subsequently leads to
secondary health benefits-efficient use of energy from food, better resorption of minerals and
vitamins by intestinal epithelium, suppression of diseases and inflammatory processes in the
human intestine and many others.(Michal Rosenberg et al., 2017)

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 CHAPTER - 3
OBJECTIVES AND METHODOLOGY

3.1. OBJECTIVES

3.1.1. To study the toxicity of cottonseed

A byproduct of the manufacture of cotton is cottonseed, which is the seed of the cotton plant
(Gossypium spp.). Cottonseed has been used historically for several uses, such as being an
industrial and culinary oil source and an element in animal feed. On the other hand, if
ingested in excessive amounts, several of the substances it contains may be hazardous to both
humans and animals. Gossypol and cyclopropenoid fatty acids are the main hazardous
ingredients in cottonseed. For example, gossypol can lead to difficulties with reproduction
and other health in animals.
1. Analyze the concentrations of harmful substances in cottonseed, such as gossypol
and cyclopropenoid fatty acids.
2. Examine the impact of these harmful substances on animal performance and health,
including growth, reproduction, and general physiological health.
3. Examine if eating meat, milk, or eggs from animals on diets high in cottonseed can
cause these toxins to bioaccumulate in animal tissues and therefore affect human
health.
4. Investigate ways to reduce or detoxify cottonseed's toxicity so that it is safer for
people or animals to eat.
5. Provide information and analysis to agricultural stakeholders, policymakers, and
regulatory bodies about the safe handling and application of cottonseed and its
derivatives.
Researchers may aid in the creation of plans to reduce hazards and guarantee the safe use of
this agricultural waste by having a better understanding of cottonseed's toxicity and possible
impacts on the health of humans and animals. This information is crucial for optimizing the

18
sustainable use of cottonseed resources across a range of sectors while preserving public
health and animal welfare.
3.1.2 Removing gossypol toxicity from cottonseed
Gossypol Reduction: The main goal is to come up with methods that will successfully bring
cottonseed's gossypol content down to levels that are safe for both human and animal
consumption. This entails investigating several physical, chemical, and biological methods to
neutralize or remove gossypol with the least amount of negative impacts on seed quality.
Preserving Nutritional Quality: It is essential to make sure that cottonseed's nutritional
content is not jeopardized by gossypol removal techniques. The goal is to create methods that
specifically target gossypol while leaving the vital nutrients—protein, lipids, vitamins, and
minerals—in the seed unaltered.
Optimization of Extraction Procedures: Research activities aim to maximize the efficiency of
gossypol removal by optimizing extraction procedures while reducing energy usage,
processing time, and environmental effects. The goal is to create scalable, affordable methods
that may be used in the industrial setting.
3.1.3 formulation of ice cream from cottonseed milk
The study aims to develop ice cream using cottonseed milk as a nutritious and sustainable
alternative to traditional dairy milk. The goal is to create a nutrient-rich product that enhances
the nutritional profile of ice cream compared to conventional dairy-based options. The
research also focuses on optimizing the formulation to achieve desirable sensory attributes,
such as taste, texture, and mouthfeel, to create a creamy, appealing ice cream that rivals or
exceeds traditional dairy ice cream. Cottonseed milk is being used in ice cream production to
create an allergen-free and vegan-friendly option, thereby expanding market accessibility and
catering to diverse dietary preferences. This also contributes to sustainability by repurposing
byproducts from the cotton industry. The aim is to create a cost-effective and scalable ice
cream using cottonseed milk, meeting consumer expectations for taste, texture, and
nutritional value. Research will assess acceptability, identify barriers, and refine the
formulation to meet market demands.
Researchers aim to showcase the feasibility and potential benefits of using cottonseed milk as
a sustainable, nutritious, and delicious alternative to traditional dairy ice cream.

3.1.4 Adding Symbiotics into the cottonseed ice cream

By integrating probiotics and prebiotics into a delectable frozen treat, symbiotics are added to
ice cream recipes to improve gut health and nutritional value. The following are this
endeavor's main objectives: The research aims to improve digestive health by developing ice
cream products with probiotic bacteria and prebiotic fibers and enhance its nutritional profile
by fortifying it with vitamins, minerals, and bioactive compounds derived from probiotics
and prebiotics.
The aim is to increase dietary fiber intake in ice cream by incorporating prebiotics, which
support digestive health. Additionally, research aims to ensure probiotic viability and stability
by selecting resistant strains and developing strategies for protection against environmental
stresses.
Symbiotic ice cream aims to balance functional benefits with sensory attributes like taste,
texture, and mouthfeel while maintaining creamy texture and flavor. Research assesses

19
product acceptability, flavor preferences, and market viability through sensory evaluations
and consumer studies.

3.2. METHODOLOGY
3.2.1 Extraction of cottonseed milk
Harvesting: Cotton plants are harvested for their seeds, which are then gathered for additional
processing. Cleaning: To get rid of any dirt, debris, or contaminants, the cotton seeds are
cleansed. Crushing: To reduce their size, the cleaned cotton seeds are crushed. Grinding: A
mill or grinder is then used to process the crushed cotton seeds into a fine powder. Mixing:
To make a thick paste, water is combined with powdered cotton seed powder. Straining: To
separate the liquid portion (cotton seed milk) from the solid particles, the paste is strained
through a small mesh or cloth. Pressing: To extract as much milk as possible, more pressure
is applied to the strained liquid. Filtering: To get rid of any last traces of contaminants, the
extracted cotton seed milk is then put through a fine filter. Pasteurization: To assure the
safety of the filtered cotton seed milk for eating, it is heated to a certain temperature, and any
hazardous bacteria are killed. Packaging: After being pasteurized, the cotton seed milk is put
into containers for distribution and storage. Quality control: Before the finished product is
made available for sale to customers, it is tested for consistency, taste, and quality.

20
 Harvesting Cleaning Crushing

Straining Mixing Grinding

Pressing Filtering Pasteurization

Quality
Packaging
control

3.2.2 Chemical analysis of cottonseed milk

3.2.2.1 Examination of fat in cottonseed milk

21
Sample Preparation: Take a sample that is indicative of the milk that will be examined.
Weighing:
Fill a Gerber fat testing vial with a precise amount of milk sample (about 10 ml).
Sulfuric Acid Addition: Fill the Gerber bottle with the milk sample and add a certain amount
of sulfuric acid (around 11 ml). Due to its extremely corrosive nature, sulfuric acid should be
handled
Centrifugation
Centrifugation involves spinning the Gerber bottle rapidly for a certain amount of time,
generally five minutes, within a centrifuge. The fat is separated from the other milk
constituents using centrifugal force.
Reading:
The fat will separate and float to the top of the milk during centrifugation. Using the Gerber
bottle's calibrated scale, determine the fat layer's thickness.
Calculation:
Use the formula or conversion sheet that came with the Gerber equipment to determine the
fat content. Usually, a percentage is used to indicate the fat content.

3.2.2.2 Examination of cottonseed milk's protein content

22
Preparing the Sample:
Get a representative sample of cottonseed milk first.
Make sure the sample is well mixed to ensure homogeneity.

Calculating the Overall Nitrogen Content:

Typically, the Kjeldahl technique is used to measure the sample's total nitrogen level to
indirectly infer the protein content. Using strong sulfuric acid, digest a specified weight of the
material (the Kjeldahl process turns nitrogen into ammonium sulfate).
Following digestion, ammonium sulfate is purified and alkalized, and the ammonia gas is
collected in a solution containing boric acid.
The nitrogen concentration is then ascertained by titrating the ammonia generated with a
standardized acid solution.

Nitrogen Conversion to Protein:

Using a conversion factor, the total nitrogen content is translated to protein content once it
has been established.
Assuming that protein includes around 16% nitrogen by weight, the conversion factor for the
majority of meals, including cottonseed milk, is normally 6.25. However, this component
may change based on the particular protein composition found in the sample.

Protein Content Calculation:

To get the protein content of the sample, multiply the total nitrogen content (derived from
step 2) by the conversion factor.
Total nitrogen content (g) × Conversion factor = Protein content (%)

3.2.2.3 Analyzing the moisture content of cottonseed milk

23
Equipment and Materials:
 Analytical balance
 Desiccator
 Weighing dishes
 Oven
 Spatula or scoop
 Cottonseed milk samples

Procedure:
Preparation of Weighing Dishes:
Clean and dry weighing dishes thoroughly.
Weigh each empty weighing dish individually and record the weights.
Sample Preparation:
To ensure homogeneity, make sure the cottonseed milk sample is thoroughly combined.
Transfer a portion of the cottonseed milk sample onto a pre-weighed weighing dish using a
scoop or spatula. Together, weigh the sample and the weighing dish, then note the total
weight.

Drying Process:
Set the oven's temperature to a desired level that will allow the sample to dry (usually
between 105°C and 110°C). Put the cottonseed milk sample-containing weighing plate in the
oven. The sample should be left to dry in the oven until its weight remains consistent.
Depending on the sample's moisture level and the drying environment, this usually takes
several hours.

Cooling and Weighing

24
The weighing dish should be taken out of the oven and allowed to cool to room temperature
in a desiccator after it has dried. Weigh the dry sample and the weighing dish together once
they have cooled. Note the ultimate weight.

Calculation
Moisture content =

3.2.2.5 Examination of water activity in cottonseed milk

The amount of water accessible for microbiological development and chemical reactions in
the sample is measured by measuring the water activity of cottonseed milk using specialist
equipment and according to prescribed procedures.

Equipment and Materials

Water activity meter
Cottonseed milk samples
Sample cups or containers
Desiccant or saturated salt solution

Procedure
Water Activity Meter Calibration:
Follow the manufacturer's instructions to calibrate the water activity meter before beginning

25
the study. To check the accuracy of the instrument, this usually entails employing saturated
salt solutions or desiccants with established water activity values.

Preparing the Sample:

To ensure homogeneity, make sure the cottonseed milk samples are thoroughly combined.
Fill a dry, clean sample cup or container with a portion of the cottonseed milk sample.
Tightly seal the sample cup to avoid losing or gaining moisture while the analysis is being
done.

Equilibration
Give the sealed sample cup with the cottonseed milk sample enough time to acclimate to the
appropriate temperature, which is typically room temperature. The sample and the particular
needs of the water activity meter might affect the equilibration time.

Measurement

Insert the sealed sample cup into the water activity meter when the sample has achieved
equilibrium. To begin the measuring procedure, adhere to the manufacturer's instructions.
The headspace air above the sample's relative humidity is measured by the water activity
meter, which then uses this information to compute the water activity.

3.2.2.6 QUALITY PARAMETERS AND MICROBIAL ANALYSIS

With possible contributory effects from direct sound waves from ultrasound equipment,
cavitation (bubbles) in the juice will induce the compression pressure and shear stress on the
microbes, causing DNA damage, and microstreaming causing thinning of cell membranes,
the antimicrobial effect of ultrasound is primarily due to the sound waves, but extreme,
pressures and temperatures produced during cavitation also leading to damage.

Total Plate Count

A microbiological method called total plate count (TPC) is used to calculate the total number
of bacteria, yeast, and molds (as well as other types of microorganisms) that are present in a
sample. It is a key technique in microbiology and is frequently employed in several sectors,
including the manufacturing of food, pharmaceuticals, and environmental monitoring

PRINCIPLE
The basic idea behind this method is that "every viable bacterium develops into a visible
colony on a nutrient agar medium when material containing bacteria is cultured." As a result,
the number of colonies equals the number of organisms in the sample. The original sample
must be diluted in order to get the desired outcome, which calls for growing, on average,
between 30-300 colonies of the target bacterium. Less than 30 colonies can result in some
flaws in explanation, whereas more than 300 colonies frequently cause overlapping and
counting mistakes.

26
RESOURCES AND APPARATUS:
Sample: The substance or material that will be tested for microbiological contamination.
Petri dishes used to cultivate microbial colonies are sterile. Agar media is a type of growth
medium that encourages the development of several microorganisms. To sterilize the agar
media, use an autoclave. Keeping a temperature under control for microbial development
(often set at 37°C for bacteria) is the purpose of an incubator. To precisely transfer and
dispense samples, use pipettes and pipette tips. A sterile solution (often buffered saline) used
to dilute the sample. To spread the diluted material on the agar plates evenly, use a sterile
spreader or glass rod. To preserve aseptic conditions, use sterile gloves. To sanitize the
spreader and workspace, use a Bunsen burner or an alcohol lamp. Labeling tape or a marker
can be used to identify plates. Record plate details and incubation conditions in the
incubation log.

PROCEDURE:
Preparation: All the tools and supplies that will come into touch with the sample should be
sterilized. This entails sterilization procedures like burning the spreader and autoclaving the
agar media. The name of the sample, the date, and any other pertinent information should be
written on the Petri dishes.

Dilution of a Sample:
To produce countable colonies, you might need to dilute the sample, depending on the
anticipated microbial load. A typical dilution technique is serial dilution, which involves
adding a known volume of the sample to a succession of sterile diluent tubes with
progressively lower concentrations. To ensure that the microbes are distributed uniformly,
vigorously mix each dilution tube.

Inoculation:
Transfer a known volume (often 1 ml) aseptically onto a sterile agar plate from the greatest
dilution tube. Spread the liquid evenly across the agar using a sterile spreader or glass rod.

Incubation:

27
 Place the inoculated agar plates in an incubator that has been adjusted to the proper
temperature for the microorganisms you are trying to count (for bacteria, this would be
37°C). Give the plates enough time to incubate (about 24-48 hours) so that colonies can
become visible

Colony Counting
Count the colonies that have grown on the agar plates after incubation. If they are simple to
recognize, count them manually or with a colony counter. Colony counts for each plate
should be noted

Calculation:
Multiplying the colony count by the dilution factor used to that plate will yield the overall
plate count. Based on the original sample, express the results as colony-forming units per
milliliter (CFU/mL) or colony-forming units per gram (CFU/g).

3.2.2.7 pH METER FOR TESTING pH

1. Calibration: First, calibrate the pH meter using a solvent-free method with a known
pH. This guarantees the measurement.
2. Preparation of liquid samples for contamination analysis. Do not forget about the
temperature.
3. Cleaning: Rinse the pH electrode with clean water to remove any remaining test.
4. Immersion in Electrode. Immerse the pH electrode in the test solution. Make sure it is
completely submerged and does not touch the sides or bottom of the container.
5. Stability: Monitor the stability of the pH reading on the instrument. This process will
take some time. Once stable, the reading should be regular. When stable, note the pH
value displayed on the digital pH meter.
6. Cleaning: Rinse the electrode again with distilled water after use.`

3.3.2.8 ACIDITY TEST

This test determines the acidity level, which is important for maintaining the flavor balance
and longevity of the beverage. To find out how acidic a beverage is, titration techniques like
titration using a standardized sodium hydroxide (NaOH) solution are commonly used. The
percentage of citric acid or another acid equivalent is used to express the results. In addition
to ensuring that the beverage satisfies required sensory qualities, such as tartness, acidity
testing provides crucial data for product stability and shelf life. The inclusion of acidity test
results in the project report for the final year shows that the chemical composition and quality
parameters of the beverage have been thoroughly analyzed and understood.

28
5.2.2.11 color analysis of cottonseed milk

Cottonseed milk may have a range of colors due to several variables including additives and
processing techniques. Like regular dairy milk, cottonseed milk often has a creamy white
appearance. However, because cottonseed proteins and oils are present, it could have a
somewhat different color.

A spectrophotometer or colorimeter can be used to precisely measure the color of cottonseed

milk. These instruments use an analysis of the light wavelengths that a material reflects or
transmits to determine its color. Typically, the color value is given as coordinates inside a
color space, such Lab* (CIELAB) or RGB (Red, Green, Blue).

It's difficult to give a precise numerical number for cottonseed milk's hue without specialized
equipment for measuring the value. It's reasonable to say, nevertheless, that the hue is a
creamy white, with some nuances according on ingredients and processing methods.

3.2.2.9 Formulation of ice cream using cottonseed milk

Get the cottonseed milk base ready. Cottonseed milk should be heated over medium heat in
a saucepan. When the sugar is all dissolved, mix it in. Add the vanilla essence, if using, and
thoroughly mix.
Let the mixture cool until it reaches room temperature.

29
3.2.2.9.1 Incorporate Symbiotic Elements:
After the combination of cottonseed milk cools down, stir in the prebiotic syrup and probiotic
yogurt. To guarantee that probiotics and prebiotics are distributed equally throughout the
mixture, thoroughly stir.

3.2.2.9.2 Cool and Freeze:

Pour the mixture into an ice cream machine and process until it becomes creamy, following
the manufacturer's directions.
Pour the mixture into a shallow container and freeze it if you don't have an ice cream
machine. Once the mixture reaches the right consistency, stir it every thirty minutes.

Ingredient
 Cottonseed milk
 4 tsp soup of sugar
 Emulsifier
 probiotic (Lactobacillus acidophilus)
 Prebiotic (Fructooligosaccharides)
 Flavors

30
Flow chart of formation of symbiotic ice cream from cottonseed milk

cottonseed milk

adding sugar mixed with an emulsifier

Blending and mixing all ingredients

Pasteurization at 72°C for 10min

Incorporating symbiotic components into ice cream

Homogenization

Cooling and aging (12hr at 4°C)

3.2.2.10 Incorporating symbiotic components into ice cream

Get the cottonseed milk base ready.
Cottonseed milk should be heated over medium heat in a saucepan. When the sugar is all
dissolved, mix it in. Add the vanilla essence, if using, and thoroughly mix. Let the mixture
cool until it reaches room temperature.

Incorporate Symbiotic Elements:

After the combination of cottonseed milk cools down, stir in the prebiotic syrup and probiotic
yogurt. To guarantee that probiotics and prebiotics are distributed equally throughout the
mixture, thoroughly stir.
Cool and Freeze:
Pour the mixture into an ice cream machine and process until it becomes creamy, following
the manufacturer's directions. Pour the mixture into a shallow container and freeze it if you
don't have an ice cream machine. Once the mixture reaches the right consistency, stir it every
thirty minutes.

31
 CHAPTER – 4

PROPOSED WORK MODULES

The study aims to create ice cream using cottonseed milk as a nutritious and sustainable
alternative to traditional dairy milk. The goal is to enhance the nutritional profile of ice cream
and optimize its formulation for sensory attributes like taste, texture, and mouthfeel. The use
of cottonseed milk in ice cream production expands market accessibility, caters to diverse
dietary preferences, and contributes to sustainability by repurposing byproducts from the
cotton industry. The study aims to create ice cream using cottonseed milk as a nutritious and
sustainable alternative to traditional dairy milk. The goal is to improve the ice cream's
nutritional profile, enhance sensory attributes, expand market accessibility, cater to diverse
dietary preferences, and contribute to sustainability by repurposing cotton industry
byproducts.

The research aims to improve gut health and nutritional value by incorporating probiotics and
prebiotics into cottonseed ice cream. The goal is to enhance the nutritional profile by
fortifying it with vitamins, minerals, and bioactive compounds derived from probiotics and
prebiotics. The research also aims to increase dietary fiber intake by incorporating prebiotics,
which support digestive health. The research also aims to ensure probiotic viability and
stability by selecting resistant strains and developing strategies for environmental stress
protection. The goal is to balance functional benefits with sensory attributes while
maintaining creamy texture and flavor.

4.1 Extraction of cottonseed milk

Cottonseed milk is a product of the extraction process of cotton plants. The process involves
harvesting the seeds, cleaning them, crushing them, grinding them into a fine powder, mixing
them with water to create a paste, straining the liquid portion, pressing more pressure to
extract more milk, filtering the milk to remove contaminants, pasteurizing it to ensure safety,
and packaging it for distribution and storage. Quality control is then conducted before the
finished product is sold to customers, ensuring consistency, taste, and quality. The process
involves several steps to ensure the safety and quality of the final product. The process
ensures the safety and quality of the final product.

4.2 The chemical analysis of cottonseed milk has been conducted.

4.2.1 Analyzing the fat content of cottonseed milk
To analyze milk fat content, take a sample of the milk to be examined. Fill a Gerber fat
testing vial with 10 ml of milk sample and add 11 ml of sulfuric acid. This is due to its
corrosive nature. Centrifugation, which spins the bottle rapidly for five minutes, separates the
fat from other milk components. The fat layer's thickness is determined using the calibrated
scale on the bottle's scale. The fat content is determined using the formula or conversion
sheet provided with the Gerber equipment, usually in percentages.

32
4.2.2 The study investigates the protein content of cottonseed milk
The Kjeldahl technique is used to measure the overall nitrogen content of a sample, which
indirectly infers protein content. This involves digesting a material with sulfuric acid,
converting nitrogen into ammonium sulfate, purifying and alkalizing it, and collecting the
ammonia gas in a boric acid solution. The nitrogen concentration is then determined by
titrating the ammonia with a standardized acid solution. The total nitrogen content is
converted to protein content using a conversion factor, typically 6.25 for most meals,
including cottonseed milk. The protein content is calculated by multiplying the total nitrogen
content by the conversion factor, resulting in protein content (%).

4.2.2 Analyzing the moisture content of cottonseed milk.

This procedure involves preparing weighing dishes, transferring a cottonseed milk sample
onto a pre-weighed dish, and drying it in an oven between 105°C and 110°C. The sample is
then transferred to the oven and left to dry until its weight remains consistent. The drying
process can take several hours, depending on the sample's moisture level and the drying
environment. After drying, the weighing dish is taken out of the oven and allowed to cool to
room temperature in a desiccator. The dry sample and weighing dish are then weighed
together, and the ultimate weight is recorded. The process ensures homogeneity and accuracy
in the analysis of cottonseed milk samples.

4.2.3 This study examines the water activity in cottonseed milk.

To calibrate a pH meter, use a solvent-free method with a known pH for accurate
measurement. Prepare liquid samples for contamination analysis, ensuring temperature is
maintained. Clean the pH electrode with clean water, then immerse it in the test solution,
ensuring it is completely submerged. Monitor the stability of the pH reading on the
instrument, recording the value displayed on the digital pH meter. After use, rinse the
electrode with distilled water.

4.2.4 ACIDITY TEST

Acidity testing is a crucial process for determining the acidity level of a beverage, which is
essential for maintaining flavor balance and longevity. It is typically done using titration
techniques, such as standardized sodium hydroxide (NaOH) solution, and is expressed as a
percentage of citric acid or another acid equivalent. This test ensures the beverage meets
sensory requirements, provides data for product stability, and ensures the quality parameters
of the beverage are thoroughly analyzed.

33
4.2.4 color analysis of cottonseed milk

Cottonseed milk's color can vary due to additives and processing techniques. It typically has a
creamy white appearance, but may have a different color due to the presence of proteins and
oils. To accurately measure the color of cottonseed milk, spectrophotometers or colorimeters
analyze light wavelengths and provide color values within a color space like Lab* (CIELAB)
or RGB (Red, Green, Blue).

4.3 Formulation of cottonseedmilk ice cream

To make a symbiotic ice cream, heat cottonseed milk over medium heat, dissolve sugar, add
vanilla essence, and let it cool. Incorporate prebiotic syrup and probiotic yogurt, stirring to
distribute probiotics and prebiotics equally. Pour the mixture into an ice cream machine or
freeze it. Stir every thirty minutes until the mixture reaches the desired consistency. The
ingredients include cottonseed milk, 4 tsp sugar soup, emulsifier, probiotic (Lactobacillus
acidophilus), prebiotic (Fructooligosaccharides), and flavors. Stir the mixture every thirty
minutes to ensure even distribution.

cottonseed milk

adding sugar mixed with an emulsifier

Blending and mixing all ingredients

Pasteurization at 72°C for 10min

Incorporating symbiotic components into ice cream

Homogenization

Cooling and aging (12hr at 4°C)

4.4 Include Symbiotic Components:

To make a symbiotic ice cream, heat cottonseed milk over medium heat, dissolve sugar, and
add vanilla essence. Allow it to cool to room temperature. Add prebiotic syrup and probiotic
yogurt, stirring to distribute probiotics and prebiotics evenly. Pour the mixture into an ice
cream machine and process until creamy, following manufacturer's directions. If not, freeze it
in a shallow container and stir every thirty minutes until the desired consistency is reached.

34
4.5 Packaing method of cottonseed milk ice cream

Ice cream packaging plays a crucial role in maintaining the quality and taste of the product. It
serves as a protective barrier, preventing spills or leaks, and preserving the frozen state.
Packaging also serves as a marketing tool, displaying branding, flavor information, and
nutritional content. It should be easy to open, handle, and consume. Common ice cream
packaging methods include individual portions, such as plastic tubs, cups, cones, popsicles,
and bulk packaging like cardboard cartons and plastic buckets.

Material selection for ice cream packaging includes food-grade plastics like Polypropylene
(PP) and High-density polyethylene (HDPE), paper and cardboard, foil and plastic films, and
edible cones. These materials offer durability, and affordability, and can withstand freezing
temperatures. Additionally, there is a growing trend towards using recyclable, compostable,
or biodegradablepackaging.

Portion control is another important consideration in ice cream packaging. Packaging can be
designed to encourage mindful portion sizes, and the choice depends on factors like the type
of ice cream, target audience, desired functionality, and brand identity. Overall, the choice of
packaging for ice cream depends on various factors.

4.storing method of ice cream

For optimal home storage of ice cream, set the freezer temperature to 0°F (-18°C) or below,
ensuring colder temperatures slow down ice crystal growth. Store ice cream towards the back
of the freezer, away from the door, to minimize temperature fluctuation. Keep store-bought
ice cream in its original packaging for best protection. Transfer homemade ice cream to an
airtight container to minimize air exposure and prevent freezer burn. Avoid putting warm ice
cream directly in the freezer and let it cool down at refrigerator temperature first. Commercial
facilities use industrial freezers with low temperatures to ensure long-term storage and
product quality. Cold chain management is crucial throughout the supply chain, from
manufacturing to distribution and retail stores. Limit door opening to prevent temperature
fluctuations and refreeze thawed ice cream to prevent quality decline and bacterial growth.
Label containers with the date of preparation for better inventory management.

35
 CHAPTER – 5

RESULTS AND DISCUSSIONS

5.1 Extraction of cottonseed milk

We received 100ml of cottonseed milk for 180g of cottonseed.

5.2 Chemical analysis of cottonseed milk

5.2.1 Fat Analysis Using Gerber Method
 We obtained 3.5 grams of fat for 100 ml of cottonseed milk

36
5.2.2 Protein analysis using the Kjeldahl technique
 After analysis, cottonseed milk typically contains 3.5- 4.5 grams of protein per 100
ml.

5.2.3 moisture content

37
  This means that for every 100 grams of cottonseed milk, there are only about 10-13
grams of solids

5.2.4 PH analysis
 cottonseed milk has a slightly acidic pH (around 6.2-6.5)

5.2.5 Color analysis of cottonseed milk

Color value L* a* b*

Cottonseed milk 35.52 15.52 35.83

 The above table shows the values of color parameters of cottonseed milk

38
 color value of cottonseed milk

90

80

70

60

50

40

30

20

10

0
cottonseed milk

L* a* b*

39
5.3 chemical properties of symbiotic ice cream from cottonseed milk

cottonseed milk ice cream

7
6
5
4
3
moisture
2
1 ph
0
cottonseed Fat
milk ice protein
cream for 100
ml

protein Fat ph moisture

3.4 Comparison of cow milk and cottonseed milk

Chemical property Cotton seed milk Cow milk

1) viscosity 2.8 centipoise 1.5 centipoise

2) Fat 3.8 gram 3 gram

3) Proteins 4 gram 3 gram

4) Lactose absent present

40
3.4.1 Comparison graph of cow milk and cottonseed milk

comparison
7

0
cow milk
cottonseed milk

protein fat ph mositure

 In contrast to cow's milk Cottonseed milk is free of lactose and contains a high
protein and fat content.

41
3.5 cost analysis of symbiotic ice cream from cottonseed milk

42
 CHAPTER 6

CONCLUSION

The conclusion emphasizes the promising future of symbiotic ice cream made from cotton
seed milk, as it offers consumers a delicious and nutritious treat that supports both their
taste buds and their well-being. By incorporating probiotics and prebiotics into the ice
cream, it not only provides indulgence but also potential health benefits such as improved
digestion, enhanced immune function, and overall gut health. Moreover, the use of cotton
seed milk contributes to sustainability efforts by utilizing a byproduct of the cotton industry.
Despite challenges like maintaining probiotic viability and optimizing flavor and texture, the
abstract suggests potential solutions and formulation strategies. Overall, the conclusion
underscores the innovative and positive impact of symbiotic ice cream from cotton seed
milk on both consumer health and environmental sustainability

43
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