S.NO CONTENTS PAGE.

NO

1 INTRODUCTION

2 LITERARURE REVIEW

3 EXPERIMENTAL PROCEDURE

4 RESULTS AND DISCUSSION

5 FINAL CONCLUSION

6 REFERENCE

7 APPENDIX

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 INDEX

S.NO COMPONENTS PAGE NO

1. INTRODUCTION 5

2. PREPARATION OF TAPIOCA STARCH 6-9

3. ANALYSIS OF IODINE COLOR REACTION 10

4. ANALYSIS OF MOISTURE CONTENT 11

5. ANALYSIS OF ASH CONTENT 12

6. ANALYSIS OF pH CONTENT 13

7. PREPARTION OF GLUCOSE SYRUP 14-19

8. PREPARATION OF ENZYME 20-24

9. ANALYSIS OF GLUCOSE SYRUP 25-31

10. LAB SAFETY PRECAUTIONS 32-33

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 Abstract:
Tapioca Starch: A Versatile Polysaccharide from Cassava

Tapioca starch, a near-pure carbohydrate derived from the roots of the

cassava plant (Manihot esculenta), is a globally significant ingredient with
diverse applications. Extracted through a process of grinding, washing, and
drying the cassava root pulp, it presents as a fine, white powder
characterized by its neutral flavor, odor, and ability to form clear, viscous
solutions and gels upon heating. Its inherent gluten-free nature has
propelled its widespread use in gluten-free baking and food formulations,
providing binding, textural enhancement, and moisture retention. Beyond
culinary applications, tapioca starch serves as a crucial raw material in
various industries, including adhesives, paper manufacturing, textiles,
pharmaceuticals, and the production of sweeteners and biodegradable
materials. While nutritionally lean, its unique functional properties,
including its thickening power, freeze-thaw stability, and film-forming
capabilities, make it an indispensable component in numerous food and
industrial processes, contributing significantly to both food security in
tropical regions and the global market for specialized starches.

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 CHAP
TER -01

INTRODUCTION:-
Tapioca starch, also known as tapioca flour, is a versatile and widely used food ingredient
derived from the cassava plant (Manihot esculenta). Here's a detailed introduction:
Origin and Extraction:
 Source: Tapioca starch is extracted from the starchy roots (tubers) of the cassava plant, a tropical root
vegetable native to South America.
 Process: The manufacturing process typically involves:
1. Harvesting and Washing: Mature cassava roots are harvested and thoroughly washed to remove
soil and impurities.
2. Grating/Crushing: The roots are then grated or crushed to release the starch contained within the
cells.
3. Starch Extraction: The resulting pulp is mixed with water, and the starch is separated from the
fiber and other components through sedimentation, centrifugation, or a combination of methods.
4. Washing and Refining: The extracted starch is washed several times to remove any remaining
impurities and proteins.
5. Drying: The purified starch slurry is then dried to a specific moisture content, resulting in a fine,
white powder – tapioca starch.
6. Milling and Sieving: Finally, the dried starch is milled to the desired particle size and sieved to
ensure uniformity.
Forms of Tapioca:
While "tapioca starch" and "tapioca flour" are often used interchangeably, tapioca is also
available in other forms:
 Tapioca Pearls (Boba): These are small, opaque or translucent spheres made from tapioca starch. They
are commonly used in desserts like bubble tea and puddings. They are made by mixing tapioca starch with
water to form a dough, which is then rolled and cut into small balls.

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 Tapioca Flakes (Instant Tapioca): These are pre-cooked and dried tapioca starch in a flake form. They
dissolve quickly in hot liquids and are often used to thicken fruit pies and sauces.
 Tapioca Syrup: This is a sweet syrup produced by enzymatically converting cassava root starch.
Properties of Tapioca Starch:
 Appearance: It is a fine, white to off-white powder.
 Taste and Odor: Tapioca starch has a very neutral flavor and no distinct odor, making it a versatile
ingredient that doesn't typically interfere with the taste or smell of other ingredients in a dish.
 Texture: In its dry form, it's a smooth, light powder. When cooked with liquid, it thickens to create a
clear, glossy, and somewhat gelatinous texture. Upon cooling, it can set into a soft gel.
 Gluten-Free: Naturally gluten-free, making it an excellent alternative for individuals with celiac disease
or gluten sensitivities.
 High Starch Content: Primarily composed of carbohydrates, with very little protein, fat, or fiber.
 Thickening Ability: It's a highly effective thickening agent, even at low temperatures. It swells and
absorbs liquids readily.
 Binding Properties: It can act as a binder in various food applications, helping to hold ingredients
together.
 Freeze-Thaw Stability: Generally exhibits good freeze-thaw stability compared to some other starches
like cornstarch, meaning it's less likely to become watery or separate after freezing and thawing.
 Clear Gel Formation: When cooked, it forms a relatively clear and shiny gel, which is desirable in fruit
fillings and glazes.
Uses of Tapioca Starch:
Tapioca starch has a wide range of applications in the food industry and beyond:
 Thickening Agent: In soups, sauces, gravies, stews, puddings, and pie fillings.
 Gluten-Free Baking: Used in gluten-free bread, cakes, cookies, and other baked goods to improve
texture and structure. It often provides a chewy and slightly springy texture.
 Binding Agent: In processed meats like sausages and nuggets to improve texture and moisture retention.
 Stabilizer: In dairy products like yogurt and ice cream for a smoother texture and to prevent separation.
 Crispy Coatings: Used to create crispy coatings for fried foods.
 Bubble Tea: The primary ingredient in tapioca pearls (boba).
 Desserts: Used in various desserts like puddings, jellies, and fruit fillings for its thickening and clear gel-
forming properties.
 Industrial Applications:
o Adhesives: Used in the production of glues and adhesives.
o Paper Industry: As a coating and sizing agent to improve paper strength and smoothness.
o Textile Industry: For sizing yarns to improve weaving efficiency and fabric finishing.
o Pharmaceuticals: As a binder and filler in tablets.
o Animal Feed: As a carbohydrate source and binder.
o Biodegradable Materials: Used in combination with biodegradable plastics.
o Sweetener Production: A significant raw material for producing glucose and fructose syrups.
In conclusion, tapioca starch is a valuable and versatile ingredient with unique properties that
make it essential in numerous culinary and industrial applications. Its gluten-free nature and
neutral characteristics have further increased its popularity worldwide.

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 Sources and related content
You're right to want to delve deeper into tapioca starch! Here's a more comprehensive look at
various aspects of this fascinating ingredient:
1. Nutritional Profile:
 Primarily Carbohydrates: Tapioca starch is almost pure carbohydrates, mainly in the form of starch
(amylose and amylopectin).
 Low in Other Nutrients: It contains very little protein, fat, and fiber.
 Minerals: It provides small amounts of minerals like calcium, iron, and potassium, but it's not considered
a significant source of these nutrients.
 "Empty Calories": Due to its high carbohydrate and low micronutrient content, tapioca is sometimes
referred to as providing "empty calories."
 Glycemic Index: Raw tapioca has a relatively high glycemic index (around 70), meaning it can cause a
quicker rise in blood sugar levels compared to some other starches. However, processing and cooking
methods can affect this.
 Resistant Starch: Cassava root contains resistant starch, which may offer some digestive benefits by
acting as a prebiotic and promoting beneficial gut bacteria.
Some modified tapioca starches are also high in resistant starch.
2. Modified Tapioca Starches:
To enhance its functional properties for specific applications, tapioca starch can undergo
various modifications:
 Pre-gelatinized Tapioca Starch: This starch has been cooked and then dried, making it instantly soluble
in cold water. It's used in instant puddings, sauces, and as a binder in dry mixes.
 Acid-Modified Tapioca Starch (Thin-Boiling Starch): Treated with mild acids to reduce viscosity when
hot but forms a firm gel when cooled. Useful in gum candies and coatings.
 Cross-linked Tapioca Starch: Treated to increase its resistance to heat, acid, and shear, making it
suitable for canned goods, pie fillings, and sauces that undergo harsh processing.
 Acetylated Tapioca Starch: Offers improved freeze-thaw stability and clarity, often used in frozen foods
and fruit fillings.
 Hydroxypropylated Tapioca Starch: Increases water solubility, freeze-thaw stability, and film-forming
properties. Used in various food and industrial applications.
 Oxidized Tapioca Starch: Has lower viscosity and forms clear, flexible films. Used in coatings and as a
sizing agent in textiles and paper.
3. Tapioca vs. Cassava Flour:
It's important to distinguish between tapioca starch (or flour) and cassava flour:
 Tapioca Starch/Flour: Made from the processed and dried starch extracted from the cassava root. It's
almost pure starch.
 Cassava Flour: Made by drying and grinding the entire cassava root. It contains starch, fiber, protein, and
other nutrients present in the whole root. It has a more fibrous texture and a slightly nutty flavor.
4. Culinary Uses in Detail:
 Gluten-Free Baking: Tapioca starch is a staple in gluten-free baking. It provides:
o Binding: Helps to hold ingredients together in the absence of gluten.
o Texture: Contributes to a light, airy, and sometimes slightly chewy texture. It can prevent baked
goods from becoming too dense or crumbly.
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 o Moisture: Helps retain moisture, resulting in softer baked goods.

 Thickening Agent:
o Clear Sauces and Gravies: Creates a glossy, translucent finish, unlike cornstarch which can make
sauces opaque.
o Fruit Fillings: Ideal for thickening fruit pies and desserts as it doesn't cloud the appearance.
o Quick Thickening: Thickens relatively quickly, even at lower temperatures.
o Freeze-Thaw Stability: Generally better than cornstarch in frozen and thawed dishes.
 Crispy Coatings: Used in batters for frying to create a light and crispy texture.
 Boba (Tapioca Pearls): The key ingredient in the chewy pearls of bubble tea. The texture and color of
boba can vary based on the preparation and any added ingredients like brown sugar.
 Desserts: Used in puddings, jellies, and other desserts for its thickening and gelling properties.
 Binding in Processed Foods: Helps bind ingredients in sausages, meat analogs, and other processed
foods.
5. Industrial Applications in Detail:
 Adhesives: Its ability to become sticky when wet makes it a base for various glues and adhesives,
including those used in paper and packaging.
 Paper Industry: Used as a surface sizing agent to improve the paper's printability, smoothness, and
strength. It can also be added to the pulp to increase paper strength.
 Textile Industry: Used for sizing yarns before weaving to reduce friction, prevent breakage, and improve
the weaving process. It also contributes to the finish and stiffness of fabrics.
 Pharmaceuticals: Acts as a binder and filler in tablets and capsules.
 Animal Feed: Used as a carbohydrate source and a binder in pelleted feed.
 Biodegradable Plastics: Incorporated into some biodegradable plastic formulations to enhance their
properties and reduce reliance on petroleum-based polymers.
 Sweetener Production: A significant raw material in the production of glucose and fructose syrups
through enzymatic hydrolysis.
6. Global Significance:
 Staple Food: Cassava is a vital staple crop in many tropical regions of the world, providing a significant
source of carbohydrates for millions of people.
 Economic Importance: The production and processing of cassava into tapioca starch and other products
contribute significantly to the economies of many developing countries.
 Gluten-Free Market: The increasing demand for gluten-free products has further boosted the importance
of tapioca starch globally.
7. Potential Drawbacks:
 Nutrient Density: As mentioned, it's low in essential nutrients.
 High Glycemic Index (in some forms): May not be ideal for individuals closely managing blood sugar
levels in its unprocessed forms.
 Processing: The extraction process can be energy-intensive.
 Cyanide Content in Raw Cassava: Raw cassava contains cyanogenic glucosides, which can be toxic if
not processed correctly. However, the processing into tapioca starch effectively removes these
compounds, making tapioca starch safe for consumption.

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 In summary, tapioca starch is more than just a simple thickener. Its unique properties,
versatility across culinary and industrial fields, and its role in gluten-free diets make it a
significant and globally relevant ingredient. Understanding its nuances, from its nutritional
profile to its various applications and forms, provides a deeper appreciation for this cassava-
derived product.
Sources and related content

Tapioca is a perennial woody shrub with an edible root, which grows in a tropical and
subtropical areas of the world Asia and southern Africa. It grows to about 8 feet and pink
flowers. Many varieties of this spectes – divided into two groups: sweet and butter tapioca –
are cultivated in the tropics for their starchy, tuberous roots. These can be processed into
tapioca ground to produce manioc or tapioca meal (Brazilian arrowrrot), used as animal fodder
or cooked and eaten as a vegetable.

Tapioca is the most important tropical root crop. Its starchy roots are a major source of dietary
energy for more 500 million people. It is know to be the highest producer of carbohydrates
among staple crops. According to United Nations Food and Agriculture Organization (FAO),
tapioca ranks fourth of food crops in developing countries after rice, maize and wheat. The
edible leaves are relatively rich in protein. Tapioca can be stored in the ground for several
seasons, and thereby serve as a reserve food when other crops fail. Tapioca is also increasingly
used for animal feed and in different industrial processes and products. A problem with tapioca
is the poisonous cyanides which have to be disarmed before consumption.

Tapioca is a starchy, granular foodstuff prepared from tapioca a roots and used as a vegetable,
in bread or as a thickening agent in liquid foods, notably puddings. “Tapioca” is a Brazilian
word, which translates as ‘juice of tapioca’. In processing, heat ruptures the starch grains,
converting them to small, irregular masses that are further baked into flake tapioca. A pellet
form, know as pearl tapioca, is made by forcing the moist starch through sieves. Granulated
tapioca, marked in various-sized grains.

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 CHAPTER-02
2. TAPIOCA STARCH PREPARATION
Tapioca starch was prepared following the method of Moorthy et al., (1999). Tapioca roots

were peeled, washed and disintegrated in a grating machine. The resulting tapioca mesh was

reground and mixed with water in the ratio 1:5 (w/v %). The mesh was filtered through double

layer of nylon cloth to obtain starch solution. The starch was separated from the water by

sedimentation and decantation. The tarch was set in an oven to dry. The native starch was

labeled 1.

Tapioca starch has many uses in a vegan kitchen. Because it is a plant-based ingredient, you
will find its use in a several dishes. From breakfast pancakes to cakes and cookies and

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thickening gravies and stews, tapioca starch has its uses. I’ll share below, how to make tapioca
starch?

Although you can easily find tapioca starch at any grocery store, most people prefer to make
their own at home. There are several reasons you should make it at home. For one, you can get
it in large quantities. Moreover, homemade tapioca starch is free from any preservatives and
artificial ingredients. It is 100% organic and natural. Also, it is very east to make. For someone
go likes to experiment with different things, making it at home can be a good experience.

Making tapioca starch is effortless and straight forward. Here is a step-by-step guide to making
tapioca starch at home.

Step-1: Start by washing the tapioca root to get rid of the dirt. Peel the root and cut it into
small chunks

Step-2: Place the chunks in a blender jug and pour water over in, make sure you have enough
water that the consistency of the final product is runny. You can add more water during
blending. You can also grate the large chunks, using a grater.

Step-3: Blend the chunks until you get a smooth consistency. In a large bowl, place a

finemesh strainer and cheesecloth over it. Pour the liquid over it.

Step-4: The residue left on the cheesecloth is important. It is actually used to make

Bammy. Allow the bowl with the white liquid to sit for an hour for the white starch to

separate and settle at the base of the bowl from the translucent liquid on top.

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Step-5: Gradually pour the liquid off and discard, what remains is tapioca starch. Scrape the
white residue using a spoon and place it on the baking tray.

Step-6: Let it dry until all the liquid is evaporated. You will get a dry block of tapioca starch.

Step-7: Place it in a food processor or spice grinder and grind until you get a smooth powder.

2.1 DEHYDRATE TAPIOCA STARCH

There are two ways of dehydrating the residue to get tapioca starch. These are:

• If you are looking for a quick way to dehydrate residue to get tapioca, place the breaking
tray in a dehydrator for about 12 to 20 hours at 120-degree F.
• Lastly, the quickest way of dehydrating residue is by placing the baking tray in an oven at
225-degree f for about 2 hours.

2.2 TAPIOCA STARCH USED

There are different ways you can is tapioca starch. Here are a few to name:

• It is most commonly used to press clothes to get the perfect crease. Mix it in water to
make a slurry and store it in a shower bottle. Spray it on clothes and press to get a crisp
cloth.
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 • It is most widely used to coat chicken or fish for frying. You can also create a batter out
of tapioca starch. Frying it will give you a crispy texture.
• It is also a thickening agent in stews and soups. You can mix it with water to create a
slurry. It helps to thicken up the soup and stews.
• It is also used in baking. You can make delicious dessert mocha is made using tapioca
starch.
• You can also create noodles from tapioca starch.
• It is also used for making boba.
• It is also used to make pancakes, tortillas, bread, cookies, and pudding.
There are several benefits of using tapioca starch. These benefits are because tapioca starch is
having high carbohydrates content and low protein content.

Furthermore, it is also a gluten-free flour making it safe for those who have gluten allergies.

CHEMICAL ANALYSIS

After extraction and modification of native starch, some chemical tests were performed
to confirm any improvements in quantity of modified starch.

• Iodine color reaction

• Moisture content
• Ash content
• pH
• Gelatinization temperature

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 CHAPTER-03
Analysis of iodine color reaction:
Apparatus required: -

a. Starch
b. Iodine solution
c. Dropper
d. Flask

Procedure: Take 1 gram of sample in a flask, disperse with 20 ml of distilled water and observe
the dissolving behavior in water. The starch is dissolves in water. Then add a few drops of the
iodine solution and observe the color reaction after mixing well.

Result: In the solution, the color changes from blue black. So, this sample is pure starch.

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 CHAPTER-04

Analysis of moisture Content:

Apparatus required: -

a. Clean dish

b. Oven

c. Stop watch

d. Desiccator (for cooling purpose)

e. Weighing machine

f. Starch

Procedure: The clean metal dishes were dried in an oven for 15 min at 105c, allowed to cool in

a desiccator for 10 min and weighed 2g of starch sample was put into the dish, weighed and

labeled. The dish was put in an oven maintained at 105c for 4 hours, After the dish were

removed, placed in the desiccator and allowed to cool for 15 min. The dish was removed from

desiccator and weighed as quickly as possible.

Calculations:

% moisture content =

(weight of wet sample – weight of dry sample) / (weight of wet sample) X 100

Result: % moisture content of starch = 16.5%

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 CHAPTER-05

Analysis of ash Content:

Apparatus required: -

a. Starch powder

b. Weighing balance

c. Crucible (dish)

d. Furnace

Procedure:

1. Take 1.5g of starch sample was weighed into a crucible of know weight

2. The crucible was put into a furnace temperature 900*c and the sample left to ash.

3. After ashing, the crucible was brought out of the furnace, cooled of the sample was

calculated.

Calculations:

% Ash content =

(weight of ash & crucible) / (weight of crucible & sample) – (weight of crucible & sample) - (weight of

crucible) X 100

Result: % Ash content in sample = 34.66%

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 CHAPTER-06
Analysis of pH Content:

Apparatus required:-

a. pH meter
b. Starch
c. Beaker
d. Distilled water
e. Weighing balance

Procedure:

1. Take 1.5g of starch sample

2. Add 10ml of distilled water
3. Stirred properly
4. The pH properly
5. The pH meter was inserted into the solution
6. The reading is taken

Result: pH of starch solution = 8.58

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 CHAPTER-07

PREPARATION OF GLUCOSE SYRUP

7.1 Introduction: The process of making glucose syrup from tapioca starch involves a few steps.
First, tapioca roots are harvested and processed to extract the starch. The starch is then mixed
with water and treated with enzymes or acid to break it down into smaller glucose molecules.
This process is called hydrolysis.

After hydrolysis, the resulting mixture is filtered to remove impurities. It is then concentrated
through evaporation to achieve the desired consistency and sweetness. The glucose syrup can be
further processes to adjust its DE value, which determines its sweetness level.

The final product is a versatile sweetener that is used in various food and beverage applications.
Glucose syrup derived from tapioca starch is know for its smooth texture and neutral flavor,
making it a popular choice in the food industry.

7.2 Preparation of Glucose syrup:

• The tapioca starch water content should be less than 18%

• Take 100g of tapioca starch was weighed.

• Add water into the starch (5.6 – 8.0)

• Put the Gluco-amylase into the liquid.

• Deliver the liquid to heating purpose with 100 – 108c steam temperature.

• We can get liquid glucose syrup.

• The conductivity value of final liquid is less than 20ppm.

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The acid hydrolysis method for the production of glucose has been replaced recently by

enzymatic treatment with three or four different enzymes. For the complete conversion into

high glucose syrup, the first step is the liquefaction intlo soluble, short-chain dextrins. A

3035% for 90 min. A temperature above 100c is preferred to assure the removal of liquid-

starch complexes. Initially, the amylase of Bacillus amyloliquefaciens was used but this has

been replaced by the amylase Bacillus amyloliquefaciens or Bacillus licheniformis. The DE

18 | P a g e
value of a starch- hydrolysate syrup depends on the time of incubation and the amount of

enzyme added. If the hydrolysate is used for the production of glucose.

The drawback of the x-amylases used currently is that they are not active at a pH below 5.9 at

the high temperature used. Therefore, the pH has to be adjusted from the natural pH 4.5 of the

starch slurry to pH 6 by adding NaOH. Also Ca2+ needs to be added because of the Ca2+

dependency of these enzymes. Pyrococcus furiosus has an extracellur x-amylase enzyme that

shows promising characteristics for applications in the starch industry. The enzyme is highly

the mostable in the absence of metal ions, active even at a temperature of 130c, and shows a

unique product pattern and substance specificity.

A large-scale starch processing industry has emerged since the mid-1900s. Before further

processing can take place the starch-containing part of the plants have to be processed and the

starch harvested. Besides starch, sugars, pentosans, fibres,

proteins, amino acids, and lipids are also present in the starch containing part of the plant. A

typical composition of a potato is a follows: 78% water, 3% protein and 17% starch. In 1811,

the German scientist Kirchhoff found that sweet-tasting syrup was obtained when starch-water

suspension was treated with diluted acid.

Only in 1931, Newkirk described a commercial process for the production of glucose from

starch. In this batch process, starch is mixed with water, boiled to dissolve the starch granules

and release the amylase and amylopectin into the water, and treated with acid for a certain

period depending on the degree of hydrolysis that is desired. Instead of boiling, a jet-cooker

csn be used in which starch is passed by mixing steam under pressure at 100-175c with starch

slurry. The heated starch slurry can then pass directly into a hydrolysis reactor for further

treatment. The enzyme, if not thermally inactivated, can be added to the starch slurry before it

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enters the jet-cooker. The starch granules are more extensively fragmented and dispersed in the

jet-cooker process train in the batch operation.

1. 2.

4.
3.

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 7.

7.3 USES OF GLUCOSE SYRUP

1. Sweetener: Glucose syrup in often used as a sweetener in candies, desserts, and baked

goods. It provides a smooth and consistent sweetness.

2. Thickener: Due to its viscous nature, glucose syrup is used as thickening agent in sauces,

jellies, and ice creams. It help to enhance texture and improve mouthfeel.

3. Moisturizer: In the production of certain confectionery items like marshmallows and

noughts, glucose syrup acts as humectants, helping to retain moisture and prevent them

from becoming dry.

4. Beverage production: Glucose syrup commonly used in production of beverages, such

as soft drinks and energy drinks. It can contribute to the sweetness and mouth feel of

these beverages.

5. Shelf-life Extension: Glucose syrup can help extend the shelf life of ceratin products by
acting as a preservative. It helps to inhibit microbial growth and maintain product quality.

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These are just a few examples of bow glucose syrup is utilized in the food industry. Its

versatility and functional properties make it a valuable ingredient in many food and beverage

applications.

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 CHAPTER-08

PREPARATION OF ALPHA AMYLASE ENZYME

8.1 Introduction:

Mangoes are a juicy tropical fruit that is popular in summer. They contain the digestive enzymes a

group of enzymes that break down carbs from starch (a complex carb) into sugars like glucose and

maltose. anylases The amylase enzymes in mangoes become more active as the fruit ripens. This is

why mangoes become sweeter as they start to ripen. Amylase enzymes are also made by the

pancreas and salivary glands. They help break down carbs so that they are easily absorbed by the

body.

That's why it's often recommended to chew food thoroughly before swallowing, as amylase

enzymes in saliva help break down carbs for easier digestion and absorption.

Bananas are another fruit that contains natural digestive enzymes. They contain amylases and

glucosidases, two groups of enzymes that break down complex carbs like starch into smaller and

more easily absorbed sugars. Like mangoes, these enzymes break down starch into sugars as

bananas start to ripen. That's why ripe yellow bananas are much sweeter than unripe green banana.

On top of their enzyme content, bananas are a great source of dietary fiber, which may aid digestive

health. A medium banana (118 grams) provides 3.1 grams of fiber.

A two-month study in 34 women looked at the connection between eating bananas and the growth

of healthy gut bacteria. Women who ate two bananas daily

experienced a modest, non-significant rise in healthy gut bacteria. However, they did experience

significantly less bloating .

8.2 Preparation of substrate:

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Banana peel used as substrate was obtained from fruit market and chopped into small pieces of

uniform size and stored in polythene bags at room temperature.

8.3 Solid state fermentation:

1. In 250ml conical flask banana peel is chopped into pieces (10gm) and it is heated at 90 degrees

in a heater.

2. 1ml of the inoculums is added and is incubated at 37 degree for 24 hours.

3. The cultivation of micro-organisms on moist solid supports, either on inert carriers or on

insoluble substrates that can, in addition, be used as carbon and energy source.

4. It holds tremendous potential for the production of alpha amylase.

5. The fermentation takes place in the absence or near absence of free water, thus being close to the

natural environment to which micro-organisms are adapted.

6. The growth of the bacillus subtilis on the banana peel is observed.

8.4 Extraction:

1. Phosphate buffer helps to extract the alpha amylase from the source and after centrifugation the

supernatant is used for the following parameters.

2. The absorbance values of crude enzyme extracts obtained from 24 and 48 hours bacterial

cultures in the presence of three substrates at 540nm.

8.5 Banana with Bacillus subtilis:

The alpha amylase activity at pH 6.7 showed maximum production at alpha amylase, which is

incubated at 37 degree for 24 hours at pH 7. A decrease or increase in enzyme production.

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 8.6 Parameters

8.6.1 Effect of pH of the medium on a amylase production:

pH of 10g (optimum) chopped banana peel was adjusted at different levels viz., 5.7,6.7,7.5,8.0

before inoculation and incubation for 24 hr. Alpha-amylase exhibits maximum activity at its definite

pH. The pH at which the enzyme exhibits maximum activity is called its optimum pH.

8.6.2 Effect of incubation temperature on a amylase production:

SSF media of banana peel (10gm) were inoculated (1 mL) and incubated at pH 7 under different

conditions of temperature as 7°C, 30°C, 37°C, 65°C for 24 hours.

Alpha-amylase exhibits maximum activity at its optimum temperature. The temperature at which

the enzyme exhibits maximum activity is called its optimum temperature.

8.6.3 Effect of substrate concentration on a amylase production:

Conical flasks containing different substrate levels (5-20 g) were inoculated (l mL) and

incubated for 24 hours at pH7 and 35'c. a-amylase catalyses the hydrolysis of a 1-4 glycosidic

linkages and producing reducing sugars. Reducing sugars like maltose is then coupled with

DNSA in alkaline medium .It produces an orange coloured complex. The intensity of the color

produced can be measured at

8.7 Testing of enzyme:

Objective: To determine if the organism is capable of breaking down starch into maltose

through the activity of the extra-cellular alpha-amylase enzyme.

Testing Procedure:

1. Use a sterile or a sterile loop to pick a few colonies from your pure culture plate. Streak a starch
plate in the form of a line across the width of the plate, Several culture can be tested on a single

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 agar plate, each represented by a line or the plate may be divided into four quadrants for this
purpose.

2. Incubate plate at 37c for 48 hours.

3. Add 2-3 drops of 10% of iodine solution directly onto the edge of colonies.
Wait 10-15 min and record the results.

• 8.8 Interpretation:
1. Positive test (“+”): The medium will turn dark. However, areas surrounding isolated colonies
where starch has been hydrolyzed by amylase will appear clear

2. Negative test (“-“): The medium will be colored dark, right up to the edges of isolated
colonies.

Figures: Two species are inoculated onto a starch palte and inclubated at 30c until growth is seen
(plate on left). The petri dish is then flooded with an iodine solution and photograph taken after
10 minutes (plate on right). Amylase positive species shows as clearing halo around the growth
(top line of growth). Amylase negative species does not have this clear halo (Bottom line of
growth).

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27 | P a g e
 CHAPTER-09
ANALYSIS OF GLUCOSE SYRUP

• Aim: To determine the amount of glucose by oxidation (iodine) method from given glucose

solution.

• Chemical Requirement:

1. 0.1 N (std) Na2S2O3.5H2o solution

2. 0.01 N (app) iodine (I2 solution)

3. 15% Na2CO3 solution

4. 1N HCL

5. Starch solution (freshly prepared)

• Chemical preparation calculations:

❖ Normality = (weight in grams)/(equivalent weight) X 100/(volume in (ml))

❖ Equivalent weight = (Molecular weight)/(R- factor)

9.1 0.1N (std) Na2S2O3.5H2o solution (for 250 ml Na2S2O3 solution):

Equivalent weight for Na2S2O3.5H2o = (248)-(1) = 248 0.1N =

(weight in grams)/(248) X (1000)/(250) Weight in grams = (0.1

X 248)/(4)

Weight in grams = 6.2 grams

9.2 1N HCL (for 50ml HCl solution):

Molarity of 37% HCL =

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(Percentage of hcl X density of HCL)/(Molecular wt of HCL) X 100
Density of HCL = 1.19 Molecular weight of

HCL = 36.5 M= (37X1.19) /

(36.5) X 10

M = 12

Normality = Molarity X (acidity (or) basicity)

N = 12x1
N = 12 N1 = 12

For 1N of HCL N2 = 1N

N1 V1 = N2 V2 V1 = X

X= (N2 V2) / N1 V2 = 50ml

X = (1x50) / (12) = 4.16ml X =

4.16ml

:- volume taken for 50ml of HCL solution = 4.16ml

9.3 15% Na2CO3 (for 20ml Na2CO3 solution):

(weight) / (volume %) = (mass of solution (g)) / (volume of solution (ml)) X 100 15% = (mass of

solution (g)) / (20) X 100

Mass of solute = 3grams

9.4 Starch solution (freshly prepared): Take 1gm of starch powder dissolved in a 100ml

distilled water and heated the solution few minutes and cool to the room temperature

and stored in a closed vessel.

Here 1% starch solution prepared

(weight) / (volume) % = (mass of solution (g)) / (volume of solution (ml)) X 100 X= 1/100 X

100

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X=1

:- 1% starch solution prepared

9.5 Glucose solution preparation:

Take 2 grams of glucose syrup dissolved in a 250ml standard bottom flask with distilled water.

Ensure the distilled water is make upto the mark only.

Principle: In alkali media, Glucose reacts with iodine (I2 soln.) and convert into gluconic acid by

oxidation. After completion of oxidation excess of hypoiodide liberates iodine on addition of

HCL. This liberated iodine is then titrated against

0.1N Na2S2O3.5H2o and thus the volume of iodine consumed in oxidation of glucose could be

found. Using following relationship amount of glucose in given solution can be determined.

2000ml. 1N I2 = 180 gm. Glucose ( C6H12O6 = 72+12+96)

Reaction equation:

9.5.1 I2 + Na2CO3 NaI + NaOI + CO2

9.5.2 C5H11O5CHO + NaOI C5H11O5COOH + NaI

9.5.3 NaI + NaOI + 2HCL 2 NaCl + H2 + I2

9.5.4 2Na2S2O3 + I2 Na S O6 + 2NaI

2 4

Procedure:
Case-I: Glucose solution sample:
i. Take 25ml of glucose solution in flask and 5ml of Na 2CO3 solution.
Mix the solution well.

ii. Add 25ml iodine solution with normality 0.01N. iii. Keep the sample

in dark room area for 30 minutes. iv. After 30 minutes take the sample

out.

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 v. Add 20ml HCL solution to reagent sample. vi. Add 1ml

starch solution to the reagent sample. vii. Take the sample for

titration against the Na2CO3 solution. viii. The sample changes the

color from blue black to the color less.

Then take the burette reading and note down in the table. Observation table (glucose solution
sample):

Burette Reading Consumption in ml

Final reading 4ml

Initial reading 0

Difference 4ml

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Difference a burette reading = Final reading – Initial reading

= 4 – 0 = 4ml
Hence, actually we added 25ml 0.01N iodine solution for oxidation of glucose

Actually, 25ml 0.01N iodine solution added for oxidation of 25ml dil. Glucose solution.

Case-II: For Blank sample:

i. Take 0.01N iodine solution in a conical flask.

ii. Add 1ml starch solution to the sample,

iii. Take the sample for titration against the Na2CO3 solution. iv. The sample changes the
color from blue black to the table. Observation table: (Blank sample):

Burette Reading Consumption of Na2CO3 in ml

Final reading 3.2

Initial reading 0

Difference 3.2

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 Calculations:

Section: -1 (Exact normality of iodine solution)

25ml iodine solution required volume 4ml of 0.1N Na2S2O3.H2o N1 V1 = N2

V2

N1 = N2 V2 / V1 = 0.1x25/25 = 0.1N

The normality of iodine solution is = 0.1N

Section: -2 (Actual experiment: - oxidation of glucose)

Hence actually we add 25ml 0.1N iodine solution in oxidation of glucose

Actually, 25ml of 0.1N iodine solution added for oxidation of 25ml of dil. glucose solution.
• 2000ml 1N iodine (I2) solution = 180gm glucose.
• 1000ml 1N iodine (I2) solution = 180/2 = 90gm glucose

• 1ml 0.1N iodine (I2) solution = 0.009gm glucose.

• 1ml 0.01N iodine (I2) solution = 0.0009gm glucose.

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Undesired vol. of 0.01N iodine solution =3.2ml

Used vol. of 0.01N iodine solution for oxidation of 25ml dil. Of glucose

= 4ml – 3.2ml

= 0.8ml

• 0.8ml, 0.01N iodine solution = 0.0009x0.8

= 0.00072gm (in 25ml dil. solution)

• In 100ml dil. Solution amount of glucose

= 0.00072x4
= 0.00288gm glucose

Result: Normality of iodine solution = 0.01N

Volume of 0.01N iodine solution required for oxidation of 25ml d dil. glucose = 0.8ml.

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 CHAPTER-10

LAB SAFETY PRECAUTIONS

• "Never put your face near the opening of a container that holds a chemical.

• "Never touch, taste or smell anything unless instructed by the teacher.

• "When pouring, pour over the sink.

• "Use care when working with acids and bases.

• "When mixing acids with water, pour acid into the water slowly.

• Ask the teacher about how to dispose of waste.

GENERAL SAFETY RULES:

• Listen to or read instructions carefully before attempting to do anything.

Notify your teacher if any spills or accidents occur.

• Wear safety goggles to protect your eyes from chemicals, heated materials, or things that

might be able to shatter.

• After handling chemicals, always wash your hands with soap and water.
• During lab work, keep your hands away from your face.

• Tie back long hair and roll up loose sleeves.

• Know the location of the fire extinguisher, fire blanket, eyewash station, and first

aid kit.

• Keep your work area uncluttered. Take to the lab station only what is necessary.

• It is suggested that you wear glasses rather than contact lenses.

35 | P a g e
• Never put anything into your mouth during a lab experiment.

• Clean up your lab area at the conclusion of the laboratory period.

• Never "horse around" or play practical jokes in the laboratory.

• Material Safety Data Sheet (MSDS) should be present in the laboratory (a document

that contains information on the potential hazards (health, fire, reactivity and

environmental) and how to work safely with the chemical product).

Glassware safety:

• Chipped or cracked glassware should not be used. Show it to the teacher.

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Conclusion:
Glucose syrup, derived from tapioca starch, is a versatile sweetener used in various food and

beverages products. It is made by extracting starch, from tapioca, then breaking it down into simpler

sugars using enzymes. The resulting liquid is filtered to remove impurities and then evaporated to

achieve the desired concentration. Glucose syrup serves as a source of energy and sweetness in

many food applications, such as confectionery, baked goods, and beverages. Its thickening

properties also make it useful in certain recipes. By using amylase enzyme, glucose syrup

containing over 50% maltose, or even over 70% maltose can be produced. The moisture content in

the glucose is 16.5%, and the ash content of the tapioca starch is 34.66%. and overall yield pf the

glucose syrup is the 78grams. The overall glucose syrup from tapioca starch is a widely utilized

ingredient in the food industry.

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 REFERENCES

 Academic and Scientific References

 Oluwatosin, B.O., et al. (2011)
Production of Glucose Syrup from Cassava Starch
Journal of Research in National Development, 9(1).
[Available online via African Journals Online (AJOL)]
— Describes enzymatic hydrolysis of cassava starch and the process of producing glucose syrup.
 Agi, A., et al. (2017)
Production and Characterization of Glucose Syrup from Cassava Starch Using Aspergillus niger and
Aspergillus flavus
Nigerian Journal of Technology (NIJOTECH), 36(1), 37-44.
— Discusses fungal enzymes used to hydrolyze cassava starch and includes analysis of the resulting syrup.
 Akinoso, R., & Olatunde, G. A. (2014)
Production and Quality Evaluation of Glucose Syrup from Cassava and Sweet Potato Starches
African Journal of Food, Agriculture, Nutrition and Development (AJFAND), 14(5).
— Compares glucose syrup production from different starch sources including cassava.
 Books and Book Chapters
 Moorthy, S.N. (2002)
Physicochemical and Functional Properties of Tropical Tuber Starches: A Review
Starch/Stärke, 54(12), 559–592.
— Good background on the properties of cassava starch relevant to hydrolysis and syrup production.
 Whistler, R.L., Bemiller, J.N., & Paschall, E.F. (1997)
Starch: Chemistry and Technology, 2nd Edition
Academic Press
— A foundational textbook covering starch hydrolysis (both enzymatic and acid) and its applications in
syrup production.

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