Chemistry IA Proposal

By: Elgine / 12F

1. Effect of Temperature and Time in Relation To Caffeine Content

RQ: How do different water temperatures and brewing time affect the caffeine content
extracted from brewing processes?

Independent Variable Dependent Variable

1. Water temperatures (for example: 3. Amount of caffeine extracted as a

80, 90, 100, 110, 120 degrees result from the brewing process
celsius)
2. Time used to brew the coffee (for
example: 3, 4, 5, 6, 7 minutes)

2. Relationship between pH and Rate of Reaction

RQ: How do different values of pH contribute to the rate of rusting of steel?

Independent Variable Dependent Variable

1. Solution containing different values 2. Rate of rusting of the steel

of pH (1, 4, 7, 10, 13)

3. Rate of Reaction in Lipase

RQ: How do different values of pH and temperature impact the denaturation of Lipase,
and which is most effective out of the 2 factors?

Independent Variable Dependent Variable

1. Value of temperature (30, 45, 60, 1. Time taken for Lipase to denature
75, 90 degrees celsius)
2. Value of pH (6, 7, 8, 9, 10)
 1. Rate of Reaction in Amylase
RQ: How do different values of pH and temperature impact the denaturation of amylase
in bananas, and which is most impactful out of the 2 factors?

Independent Variable(s) Dependent Variable(s) Control Variable(s)

1. Temperature of water 2. Amount of amylase 1. Amount of water used

(30, 45, 60, 75, 90
degrees celsius)
2. pH of water (6, 7, 8,
9, 10)
extracted for every trial
2. Room temperature for
every trial
3. Weight of banana
used for every trial
4. Duration in which the
banana is placed in
the water for every
trial

- How to measure amylase(titration/gas chromatography, by

measuring fatty acid content (?) )
- How to extract amylase (centrifugation, ultrafiltration,
lyophilization)
- How to measure amylase activity
Extraction of Lipase
https://biomedres.us/fulltexts/BJSTR.MS.ID.001913.php#Results%20and%20Discu
ssion

Avocado oil → Fatty acid

https://iopscience.iop.org/article/10.1088/1757-899X/725/1/012075/pdf

Extraction of Lipase Enzyme from Avocado Seeds

http://jurnal.kimia.fmipa.unmul.ac.id/index.php/JA/article/view/486/321

Extraction of Lipase Enzyme from Sunflower Seeds

https://www.tandfonline.com/doi/abs/10.1081/PB-200041442?journalCode=lpbb20

Purification method of glycerol molecule

https://www.frontiersin.org/articles/10.3389/fchem.2019.00774/full

Method of extracting enzyme

https://www.google.com/url?sa=t&source=web&rct=j&url=https://www.scielo.br/j/
babt/a/sqJsWgp9PVL9XN3k7R6yT5G/%3Flang%3Den&ved=2ahUKEwjb-rLZ4vr6A
hXmaGwGHYvnAmsQFnoECA0QBQ&usg=AOvVaw2JjlfpEQOIlkGe_VGLk3k-

How to measure amylase activity

https://www.sigmaaldrich.com/ID/en/technical-documents/technical-article/protein
-biology/enzyme-activity-assays/amylase-activity-assay-kit-mak009

Amount of Banana Wasted Everyday

https://www.epicurious.com/archive/blogs/editor/2014/03/bananas-are-thrown-out
-every-day-recipes-and-tips.html

Production of Banana in 2020

https://www.statista.com/statistics/716037/global-banana-market-volume/

How temperature reduces enzyme activity

https://www.bbc.co.uk/bitesize/guides/z9jrng8/revision/3#:~:text=How%20tempera
ture%20affects%20enzyme%20action,enzyme%20will%20have%20been%20denat
ured%20.

How pH affects enzyme activity

https://byjus.com/question-answer/how-does-ph-affect-enzyme-activity/
Impact of amylase in bananas
https://www.ijcmas.com/vol-2-10/Moiedi%20Sunira%20Paul%20and%20V.%20Judi
a%20Harriet%20Sumathy.pdf

Journal for Extracting Amylase from Dragon Fruit Peels

https://www.hindawi.com/journals/tswj/2014/640949/#methods-and-materials
 Session: May 2022

International Baccalaureate Diploma Programme

Chemistry (Standard Level)

Internal Assessment

Research Question: How do different values of pH and temperature impact the

denaturation of amylase in bananas?

Word count: xxxx words

 Table of Contents

I. Introduction 1

II. Exploration 2
2.1 Background Information 2
2.1.1 Denaturation of Enzyme 2
2.1.2 Amylase Content in Banana 2
2.2 Research Question 2
2.3 Hypothesis 3
2.4 Variables 3
2.5 Tools and Materials 4
2.6 Methodology 4
2.6.1 Preparation of Banana 5
2.6.2 Preparation of Water with Different pH Levels 5
2.6.3 Preparation of Water with Different Temperatures 5
2.6.4 Amylase Activity Assay 6

III. Findings 6
3.1 Data 6
3.2 Graphs 6
3.3 Analysis 6
I. Introduction

Bananas, often grown in tropical countries, are one of the most produced fruit in the
world. According to Statista.com, 119.83 million tons of bananas are produced in the year
of 2020. However, banana is also a fruit which produces the most wastes with 60% of its
biomass being left as a waste during its harvest. As a basketball athlete, me and my team
were often given bananas before competitions as it is a good source of energy. Yet, there
are always a bunch of leftovers. This made me wonder about how much bananas are
actually being wasted around the world. Turns out, more or less, there are 10 million
hectares of banana plantation worldwide with each generating 220 tonnes of waste every
year. Furthermore, humans as consumers themselves throw away a considerable amount
of bananas, approximately 1.4 million everyday. The worst part is, these agri-wastes
release methane which is a greenhouse gas that can negatively impact the environment.

Fortunately, bananas which are wasted are not entirely useless and can become beneficial.
Bananas contain enzymes known as amylase which is a protein that helps in digestion.
They are also often used in industries for baking, brewing, fermenting, and even
pharmaceutical purposes. Hence, amylase can be extracted from the wasted bananas,
making it useful for further industrial purposes. However, there may be several factors
that can affect the amylase content of the wasted bananas. Meanwhile, most industries
would like their processes to be as efficient as possible. Therefore, this paper will aim to
examine how factors such as the different values of pH and temperature impact the
denaturation of amylase in bananas. This way, the optimum condition for effectively
extracting amylase from bananas can be discovered.

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II. Exploration

2.1 Background Information

2.1.1 Denaturation of Enzyme
“An enzyme is a biological catalyst that speeds up the rate of a chemical reaction
and is almost always a protein” (National Human Genome Research Institute, 2022).
Enzymes are mainly found in the human digestive system such as the liver, pancreas,
stomach, etc. They can also be found in other sources such as fruits.

However, there are 2 main factors that can denature enzymes which are
temperature and pH level. The definition of denaturation itself is the disruption process of
the structure of enzymes and proteins, resulting in the degrading of the cell’s activity. At
lower temperatures, the kinetic energy of the molecules in a cell decreases, leading to less
successful collisions and lower enzyme activity. On the other hand, higher temperatures
can alter the shape of an enzyme’s active site. Hence, reducing its activity or even
causing it to stop working.

The pH level can also affect the activity of an enzyme. When not under its
optimum pH level, the ionization between amino acids and atoms may take place,
resulting in the transformation of the protein’s structure.

2.1.2 Amylase Content in Banana

As mentioned previously, enzymes are commonly found in the human body. One
of those enzymes is amylase which is mainly secreted by the pancreas and salivary
glands. It functions to convert starch and glycogen into simple sugars. However, it is also
widely used for industrial purposes all around the world, especially for biotechnology and
medical practices.

Studies have proven that amylase are also found in banana pulps. In fact, it is
what causes bananas to ripen and taste sweet (M.S Paul, 2013). This is the main reason as
to why wasted bananas can be processed to become beneficial for industrial uses. Add
theoretical amylase content of a banana!

2.2 Research Question

How do different values of pH and temperature impact the denaturation of
amylase in bananas?

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 2.3 Hypothesis
Hypothetically, a pH and temperature that is either too low or too high will
increase the denaturation of amylase in bananas. A pH that is acidic (too low) or basic
(too high) will slow down the enzyme activity by affecting the bonds which contain
charges. Similarly, extreme temperatures will disrupt its active sites and slow down its
activity, denaturing the enzyme. Therefore, a neutral pH level and room temperature will
be the optimum conditions for preventing the denaturation of amylase in bananas.

2.4 Variables
Independent variable: Temperature and pH level of the water in which the bananas are
placed in.
Dependent variable: Amylase content/activity inside the bananas.

Controlled Variable Purpose Method of Controlling

1. Weight of bananas To ensure that the potential
used. amylase content of the
bananas used in every trial is
equal.
Before every trial, the weight
of the bananas are measured
with a weighing scale until it
is according to the
predetermined weight.. This
way, equal masses of bananas
can be used for every trial.

2. Duration of the So that the bananas in every
bananas placed inside trial undergo the same
the water. treatment conditions. This
way, the rate of denaturation
in every trial is equal.
When performing each trial, a
stopwatch must be used to
keep track of the duration.
Once the stopwatch has
reached the predetermined
duration, the bananas must
immediately be removed
from the solution to prevent
further denaturation.

3. Amount of water To ensure that the volume and
used. amount of molecule in which
the bananas are placed are
equal.
Prior to preparing the solution
in which the bananas are
going to be placed in, the
volume of water must be
measured with volumetric
flasks. It is imperative that
each trial should have the

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 same volume of water
measured in the volumetric
flasks.

4. The room temperature To prevent external factors The trials must be done in the
in which the from influencing the same room. Furthermore, the
experiment is condition of the internal room must be a closed room
conducted. system. This way, a more with tools such as an air
accurate experimentation conditioner to maintain a
result can be extracted. steady room temperature.

2.5 Tools and Materials

Materials:
- Bananas
- Distilled water
- Tap water
- Baking soda
- Hydrochloric acid
- Cups
- Beakers
- Containers
- Plastic wraps
- DNS
- Sodium acetate buffer
- Soluble starch
- Maltose

Tools:
- Knife
- Volumetric flask
- Water bath
- Blender
- Spectrophotometer

2.6 Methodology
Add 5 trials for untreated bananas to measure the maximum potential amylase content (?)

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2.6.1 Preparation of Banana
1. Cut several bananas into slices of similar size.
2. Blend the slices of banana using a blender.
3. Weigh the blended bananas until they are 100 g and put them into a plastic cup.
4. Repeat steps 1-3 until there are a total of 30 cups with 100 g of bananas each.

2.6.2 Preparation of Water with Different pH Levels

1. Measure 300 mL of distilled water with a volumetric flask and pour it into a
beaker.
2. Repeat step 1 until there are 15 different beakers with 300 mL of water in each.
3. Pour hydrochloric acid into a beaker to lower the pH value.
4. Repeat step 3 until there are 3 beakers with a pH value of 2, 3 beakers with a pH
value of 4, and 3 beakers with a pH value of 6.
5. Pour baking soda into the remaining beakers.
6. Repeat step 5 until there are 3 beakers with a pH value of 8 and 3 beakers with a
pH value of 10.
7. Put the 100 g of bananas into all 15 different beakers.
8. Cover the lid of the beakers with a plastic wrap and let it sit for 60 minutes.
9. After 60 minutes, extract the bananas from the beakers and put them back into the
plastic cups.
10. Cover the 15 plastic cups with a plastic wrap and store them 15 in a container
under room temperature.
11. Add homogenization of buffer and centrifugation of filtrate (?)

2.6.3 Preparation of Water with Different Temperatures

1. Measure 300 mL of distilled water with a volumetric flask and pour it into a
beaker.
2. Repeat step 1 until there are 15 different beakers with 300 mL of water in each.
3. Out of the 15 beakers, take 3 beakers and put them into a water bath.
4. Set the temperature of the water bath to 30 degrees celsius.
5. Once the temperature has been reached, put the 100 g of bananas into the 3
different beakers.
6. Let it incubate in the water bath for 60 minutes.
7. After 60 minutes, take out the beakers from the water bath, move the bananas into
the plastic cups, and cover them with a plastic wrap.
8. Store the 3 wrapped plastic cups in a container under room temperature.
9. Repeat steps 1-8 but change the temperature of the water bath to 45, 60, 75, and
90 degrees celsius instead. At the end, there should be 15 wrapped plastic cups
stored in a container under room temperature.

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 2.6.4 Amylase Activity Assay
1. Create a mixture by pouring 100 g of 0.1% soluble starch into a 0.1 M sodium
acetate buffer at pH 5.0.
2. Mix the mixture with a cup that contains the 100 g of treated bananas.
3. Incubate the mixture under 70 degrees celsius for 30 minutes.
4. After 30 minutes, add 100 g of DNS into the mixture and continue heating it for 5
minutes in boiling water.
5. After 5 minutes, cool the mixture with tap water.
6. Using a spectrophotometer at 540 nm, measure the released reducing sugar with
maltose as a standard reducing sugar. One unit of amylase activity is equivalent to
1 micromol of maltose produced per minute under the above conditions.
7. Repeat steps 1-7 for the remaining 29 cups that contain 100 g of treated bananas.

III. Findings

3.1 Data
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3.2 Graphs
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3.3 Analysis
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