Khan

ABSTRACT

The Illinois Junior Academy of Science

This form/paper may not be taken without IJAS authorization.

CATEGORY Pharmaceutical STATE REGION # 6

SCHOOL Niles North High School IJAS SCHOOL # 6038
CITY/ZIP Skokie, 60077 SPONSOR CELL PHONE #
Constantine
SPONSOR Theodoropoulos SPONSOR E-MAIL conthe@d219.org

MARK ONE: EXPERIMENTAL INVESTIGATION ☐ DESIGN INVESTIGATION ☐

NAME OF SCIENTIST* Hasan Khan GRADE 11

NAME OF SCIENTIST GRADE
NAME OF SCIENTIST GRADE
NAME OF SCIENTIST GRADE

* If this project is awarded a monetary prize, the check will be written in this scientist's name, and it will be his/her responsibility to distribute the
prize money equally among all participating scientists.

PROJECT TITLE Effects of Melatonin, Caffeine, and Nicotine on Daphnia Heart Rate

Purpose:

The purpose of this research is to measure the effects these common substances have on the
heart of Daphnia. Caffeine, Melatonin, and nicotine are all very known and used substances.
However the effects of these are not given much attention. This project helped figure out how
each substance affects the heart rate of Daphnia.

Procedure:

For this experiment, Daphnia were housed in one container where they would live. At time of
testing, daphnia would be taken by pipette and put into a beaker with the solution of the
substance for ten minutes. One these ten minutes pass, the Daphnia are placed onto a
microscope slide where they would be measured. After taking trails of the Heart Rate, Daphnia
were put into 3 separate containers. One for caffeine, one for melatonin, and another for
nicotine. Daphnia would continue to live here to compare how long each Daphnia survived.
Conclusion:

When reflecting upon the data, we see that caffeine had the lowest heart rate, melatonin had the
second highest, and nicotine had the highest with a total of 476 bpm. This is not what was
expected and was quite a shock. It was found that melatonin and nicotine both accounted for a
spike in heart rate compared to caffeine.

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SAFETY SHEET
The Illinois Junior Academy of Science

Directions: The student is asked to read these introductions carefully and fill out the bottom of this sheet. The
cience teacher and/or advisor must sign in the indicated space. By signing this sheet, the sponsor
ssumes all responsibilities related to this project.

afety and the Student: Experimentation or design may involve an element of risk or injury to the student, test
ubjects and to others. Recognition of such hazards and provision for adequate control measures are joint
esponsibilities of the student and the sponsor. Some of the more common risks encountered in research are
hose of electrical shock, infection from pathogenic organisms, uncontrolled reactions of incompatible
hemicals, eye injury from materials or procedures, and fire in apparatus or work area. Countering these
azards and others with suitable safety practices is an integral part of good scientific research. In the chart below,
st the principal hazards associated with your project, if any, and what specific precautions
ou have used as safeguards. Be sure to read the entire section in the Policy and Procedure Manual of the Illinois Junior Academy of Scienc
Guidelines for Experimentation" before completing this form.

ossible hazards Precautions taken to deal with each hazard

1. Exposure to Melatonin, Caffeine, and Nicotine 1. Work with gloves and safety goggles on
2. Exposure of chemicals to the lab 2. Work cautiously in the fume hood
3. Contact with skin 3. Frequently wash hands and materials

pecific safety practices related to materials requiring endorsement sheets should be detailed on the specific
ndorsement sheet and not included on this safety sheet.

lease check off any other possible endorsements needed. Include these documents in your paper and on your board.
____Humans as Test Subjects –for any projects involving humans including survey administration;
____Microorganisms-for any projects involving bacteria, viruses, yeasts, fungi or protazoa;
____Non-Human Vertebrates -for any projects involving fish, amphibians, reptiles, birds or mammals;
____Tissue Culture-for any projects involving growing eukaryotic tissues or cell cultures;
____Letter from institution where research was done or IJAS SRC, if an exception to the IJAS rules has been granted…

IGNED
Hasan Khan
Student Exhibitor(s)
IGNED
Sponsor *
*As a sponsor, I assume all responsibilities related to this project.

his Sheet Must Be Typed and This form must be displayed on the front of the exhibitor’s display board. It may be
educed to half a sheet of paper 8.5 inches (vertical) X 5.5 inches (horizontal).

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Effects of Melatonin, Caffeine,

and Nicotine on Daphnia Heart
Rate

Hasan Khan
Niles North High School

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Table of Contents
Acknowledgments…………………………………………………………5
Purpose & Hypothesis……………………………………………………6
Review of Literature………………………………………………………7
Materials………………………………………………………………11
Methods and Procedures………………………………………………12
Variables…………………………………………………………………13
Results
Data…………………………………………………………………14
Data Analysis………………………………………………………15
Conclusion…………………………………………………………………16
Reference List…………………………………………………………….17

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Acknowledgements
A special thanks to my family, especially my sister. My sister took this same class

when she went to Niles North and she was the one that informed me about the class

and encouraged me to take it. My family was supportive throughout the entire process

and always was trying to help. When I decided I wouldn’t do science fair they were

supportive of my choice and understanding.

This project would also be impossible without the help of my peers as well. All of my

classmates made my experience very memorable and enjoyable. They were always

willing to help out and made the class better in every way.

Last but not least, I would like to thank my teacher, Mr. Theodoropoulos. Mr. T was

always there to help me and my peers out and was very patient with all of us. Aside

from all the help he gave during the project itself, he made the class fun and kept it

balanced throughout the year.

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Purpose
The purpose of this project was to study the effects that melatonin, caffeine, and

nicotine can have on your heart rate. These three substances are known world wide

and are used by many everyday. I believed that it was important to understand what

these do to one of your major organs, your heart. The experiments aim was to see the

increase or decrease in heart rate.

Hypothesis
When exposed to melatonin, caffeine, and nicotine, Daphnia heart rates will either

increase or decrease.

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Review of Literature

Have you ever wondered why you feel a sudden burst of energy as soon as you

drink coffee, or why you get drowsy so fast when you eat melatonin? This has much

more to do with your heart than you’d think. In this pharmaceutical research, the change

in heart rate levels for Daphnia in response to melatonin, caffeine, and nicotine will be

measured. Separate colonies of Daphnia will be in different environments. There will be

three colonies, each containing either melatonin, caffeine, or nicotine. The heart rates of

each daphnia group will be recorded 10 minutes after they have been placed into a

certain tank. Daphnia are a suitable organism to use as they are incredible model

organisms for pharmaceutical research due to their physical traits.

Melatonin and caffeine are used by us humans for opposite reasons. We use

melatonin to increase melatonin production which allows for falling asleep faster and

falling asleep at different times than usual. Caffeine, as we all know, is mainly found in

coffee and soda. By consuming caffeine, we feel more energized and awake throughout

the day. That is why a lot of us need a cup or two of coffee to get us through our day.

Nicotine is also one of the most known stimulants. It is highly addictive and harmful, with

major harmful effects on your heart. The following paragraphs will give more detail

regarding the organism and substances that make this research whole.

This research project revolves around daphnia. Daphnia are known as crustaceans

and belong to the Branchiopoda group, and they fall into the Cladocera category

(Bethesda, 2005).

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Daphnia can be found all over the world. They inhabit almost all types of freshwater

bodies with the exception of extreme circumstances, such as hot springs (Bethesda,

2005).

Daphnia is a very simple model organism when it comes to studying the heart.

Daphnia has a clearish layer of skin, making it basically transparent. This allows for an

accurate and precise measurement of its heart rate and other organs. Daphnia are the

perfect model organisms for testings like this. These beneficial aspects are a reason

why it is one of the most used organisms in pharmaceutical research (Tkaczyk et al.,

2021).

The biggest advantage of using Daphnia, for this specific research, is the ability to

immobilize them very easily (Tkaczyk et al., 2021).

This can be done by having a tiny amount of water in the microscope slide as it will

decrease the space for the Daphnia to move and allow us to examine their heart rate.

One of the main aspects that makes this study whole, is the use and understanding

of melatonin. In this experiment melatonin will be one of the things being tested on the

daphnia magna. Melatonin is something that our body already produces. The production

of melatonin takes place in the pineal gland, located right above the thalamus (Claustrat

& Leston, 2015). When you are in the dark at a routine time, your brain starts to produce

natural melatonin, which is why we fall asleep in the dark (Claustrat & Leston, 2015).

However, you can also find melatonin at a local drug store. This is to aid you when

struggling to fall asleep. In regards to the research, melatonin is expected and known to

reduce heart rates in animals and humans (Simko et al,. 2016).

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The effects of melatonin will be compared to the effects that other substances have,

such as nicotine and caffeine.

Whenever something good happens and you feel happy, that’s the dopamine being

released from your substantia nigra, ventral tegmental area, and hypothalamus of the

brain. Now what does this have to do with your heart rate (Olguín et al,. 2015)?

Dopamine was found to cause an increase in your pulse pressure, and heart rate.

However dopamine is something that is only made when you are feeling happy because

of something. Yet there is still a certain stimulant that you and three quarters of America

probably drank this morning that could have led to the production of dopamine; coffee.

Caffeine is known to increase dopamine production in your brain (Volkow et al,. 2015).

Too much dopamine can increase the heart rate to a higher than normal level which

is not desirable. Having a resting heart rate over 100 beats per minute can also lead to

sinus tachycardia (Olshansky & Sullivan, 2019).

Sinus tachycardia is a clinical syndrome that can cause palpitations, discomfort, and

lead to more problems (Olshansky & Sullivan, 2019).

By having a single daphnia in a small amount of caffeine infused water, we can test

to see what the effects of caffeine are on heart rate. Since it is already known that

caffeine and dopamine work together and caffeine consumption leads to increased

dopamine production, we can expect to see an almost immediate increase in the

daphnias activity and heart rate levels as well(Kundu & Singh, 2018).

Nicotine has been known for many years to be incredibly harmful to your body. The

harm caused from nicotine varies from harming your heart, reproductive system, lungs,

kidneys, and much more (Mishra et al,. 2015).

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With its known lethal effects, what effect does it have on your heart rate? It was

discovered that ~1 mg of nicotine was enough to increase heart rate in young healthy

adults (Jensen et al,. 2020).

This can also cause sinus tachycardia just like caffeine. For the experiment

powdered nicotine will be infused into the tub of water. Once 10 minutes pass, the

daphnia will be placed under the microscope and its heart rate will be recorded. With

cigarette use being the number one preventable killer in America, there are still 50.6

million nicotine consumers whether it is electric or a cigarette (Cornelius et al,. 2020).

That means there are 50.6 million healthy adults ruining their health due to the

nicotine in these products. There are countless dangers to an elevated heart rate. By

taking this addictive stimulant, your heart will weaken and many other organs won’t be

able to function properly in the long run.

Now that we have concluded our research, we can conclude that melatonin is

beneficial for the decrease of heart rate, caffeine is responsible for increase in heart

rate, and nicotine also increases heart rate. There can be dangers associated with the

overconsumption of all three substances. To truly understand the effects on heart rate,

daphnia will be effective organisms as their translucent skin will allow us to observe the

changes in heart rate. From the results of the study, we will gain an increased

understanding on how these common substances affect our heart rate and it will be

supported with a visual representation. This research should inform people on why it is

important to be conscious of our diet as it can directly affect our heart and motivate

individuals to practice keeping our heart rate at a steady and normal rate.

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Materials
● 90 Daphnia

● 25 grams of melatonin

● 6 clear plastic containers

● 225 gram powdered spirulina

● 100 g caffeine

● 6 gallon spring water

● 100 ml nicotine

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Procedure
1. Set up 3 containers with 3.785 liter (1 gal)of water, keep equal amount of daphnia

in each. Daphnia will live here and be fed here with dosage of 1 gram powdered

spirulina

2. Set up 3 more containers, gallon each., Infuse with 0.001g melatonin, one with

0.001g caffeine, and one with 0.001g nicotine

3. At time of testing, take daphnia and place in controlled environment for 10 minutes

before examining

4. Prepare a slide with a couple drops of water to count the daphnia's heart rate

5. Place back into the pure water environment and let them recover

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Variables

Independent variable: Melatonin, caffeine, and nicotine

Dependent: Heart rate of the daphnia

Controlled variable: Time of exposure, grams of substances

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Data

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Data Analysis
In the first graph, the measurements of daphnia heart rate right after exposure is shown.

We can infer that nicotine had the highest heart rate of all, melatonin had the second

highest, base was the third highest, and caffeine resulted in the lowest heart rate of them

all.

The second graph shows the heart rates of daphnia after four days have passed. One

obvious trend here is that all the daphnia's heart rates dropped down after four days.

Nicotine had the highest, caffeine had the second highest, and melatonin had the lowest

this time. From the two graphs we can infer that melatonin spikes up and then drops back

down over time.

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Conclusion
The main purpose of this research project was to see how caffeine, melatonin, nicotine

affect the heart rates of daphnia. It was assumed that caffeine would result in the highest

heart rate, followed by nicotine, and then melatonin. However, post research we can look

back at the data and say that this was not the expected result. We can conclude that

caffeine had the lowest heart rate, then melatonin, and nicotine with the highest.

This study alters into our personal lives a lot. A lot of us are dependent on caffeine. By

starting off your day with a cup of coffee, you are essentially causing your heart rate to

decrease. This can have its own complications and problems associated with it. Many are

also reliant on melatonin to fall asleep. However, not many know that melatonin can cause

an increase in heart rate which can also lead to complications. Lastly, nicotine, so many

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people are addicted to such a harmful substance. Consuming nicotine can also lead to

increased heart rate which can lead to problems as severe as heart attacks, and

palpitations.

We should all be cautious of what we put into our bodies. Proved by this research,

effects like these can cause harm in the long run.

Work Cited
Simko, F., Baka, T., Paulis, L., & Reiter, R. J. (2016). Elevated heart rate and nondipping

heart rate as potential targets for melatonin: a review. Journal of pineal research, 61(2),

127–137. https://doi.org/10.1111/jpi.12348

Kundu, A., & Singh, G. (2018). Dopamine synergizes with caffeine to increase the heart

rate of Daphnia. F1000Research, 7, 254.

https://doi.org/10.12688/f1000research.12180.1

Tkaczyk, A., Bownik, A., Dudka, J., Kowal, K., & Ślaska, B. (2021). Daphnia magna

model in the toxicity assessment of pharmaceuticals: A review. The Science of the total

environment, 763, 143038. https://doi.org/10.1016/j.scitotenv.2020.143038

Claustrat, B., & Leston, J. (2015). Melatonin: Physiological effects in humans.

Neuro-Chirurgie, 61(2-3), 77–84. https://doi.org/10.1016/j.neuchi.2015.03.002

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Juárez Olguín, H., Calderón Guzmán, D., Hernández García, E., & Barragán Mejía, G.

(2016). The Role of Dopamine and Its Dysfunction as a Consequence of Oxidative

Stress. Oxidative medicine and cellular longevity, 2016, 9730467.

https://doi.org/10.1155/2016/9730467

Ziegler, M. G., Kennedy, B., Holland, O. B., Murphy, D., & Lake, C. R. (1985). The

effects of dopamine agonists on human cardiovascular and sympathetic nervous

systems. International journal of clinical pharmacology, therapy, and toxicology, 23(4),

175–179.

Mishra, A., Chaturvedi, P., Datta, S., Sinukumar, S., Joshi, P., & Garg, A. (2015).

Harmful effects of nicotine. Indian journal of medical and paediatric oncology : official

journal of Indian Society of Medical & Paediatric Oncology, 36(1), 24–31.

https://doi.org/10.4103/0971-5851.151771

Cornelius, M. E., Wang, T. W., Jamal, A., Loretan, C. G., & Neff, L. J. (2020). Tobacco

Product Use Among Adults - United States, 2019. MMWR. Morbidity and mortality

weekly report, 69(46), 1736–1742. https://doi.org/10.15585/mmwr.mm6946a4

Olshansky, B., & Sullivan, R. M. (2019). Inappropriate sinus tachycardia. Europace :

European pacing, arrhythmias, and cardiac electrophysiology : journal of the working

groups on cardiac pacing, arrhythmias, and cardiac cellular electrophysiology of the

European Society of Cardiology, 21(2), 194–207.

https://doi.org/10.1093/europace/euy128

Cornelius, M. E., Wang, T. W., Jamal, A., Loretan, C. G., & Neff, L. J. (2020). Tobacco

Product Use Among Adults - United States, 2019. MMWR. Morbidity and mortality

weekly report, 69(46), 1736–1742. https://doi.org/10.15585/mmwr.mm6946a4

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Ebert D. Ecology, Epidemiology, and Evolution of Parasitism in Daphnia [Internet].

Bethesda (MD): National Center for Biotechnology Information (US); 2005. Chapter 2,

Introduction to Daphnia Biology. Available from:

https://www.ncbi.nlm.nih.gov/books/NBK2042/?report=classic

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