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Name : Ahmad Luqman Bin Md Asri
IC Number : 950202146067
Group : 11SC1
SID Number : 1311170102
Title : The effect of caffeine on heart rate.
Date of Experiment : 2/10/2013
Date of Submission : 9/10/2013
Lecturer: : Puan Lili Syahani Binti Rusli


The effect of caffeine on heart rate.
1. To investigate the effect of caffeine on the heart rate of daphnia.
2. To highlight experimental and investigative skills.
This experiment is conducted to study the relationship between caffeine concentration
and the heart rate. It is carried out to study whether the heart rate is affected by different
concentration of caffeine. For this experiment, daphnia sp was used. as the experimental
materials as Daphnia sp. are the easiest species to be seen under light microscope.
Daphnia is a genus of small, planktonic crustaceans, between 0.2 and 5 mm in length.
Daphnia are members of the order Cladocera, and are one of the several small aquatic
crustaceans commonly called water fleas because of their saltatory swimming style (although
fleas are insects and thus only very distantly related). They live in various aquatic environments
ranging from acidic swamps to freshwater lakes, ponds, streams and rivers.
The two most readily available species of Daphnia are D. pulex (small and most
common) and D. magna (large). They are often associated with a related genus in the order
Cladocera: Moina, which is in the Moinidae family instead of Daphniidae and is much smaller
than D. pulex (approximately half the maximum length). Daphnia eggs for sale are generally
enclosed in ephippia (a thick shell, consisting of two chitinous plates, that encloses and protects
the winter eggs of a cladoceran.)
Sometimes Daphnia may be used in certain environments to test the effects of toxins on
an ecosystem, which makes them an indicator genus, particularly useful because of its short
lifespan and reproductive capabilities. Because they are nearly transparent, their internal organs
are easy to study in live specimens (an example might be to study the effect of temperature on
the heart rate of these ectothermic organisms). They are often fed to tadpoles or small species of

amphibians such as the African dwarf frog (Hymenochirus boettgeri). Daphnia spp. are also a
popular live food in tropical and marine fish keeping.
Caffeine is a bitter, white crystalline xanthine alkaloid and a stimulant drug. Caffeine is
found in varying quantities in the seeds, leaves, and fruit of some plants, where it acts as a
natural pesticide that paralyzes and kills certain insects feeding on the plants, as well as
enhancing the reward memory of pollinators. It is most commonly consumed by humans in
infusions extracted from the seed of the coffee plant and the leaves of the tea bush, as well as
from various foods and drinks containing products derived from the kola nut. Other sources
include yerba mat, guarana berries, guayusa, and the yaupon holly.
In humans, caffeine acts as a central nervous system stimulant, temporarily warding off
drowsiness and restoring alertness. It is the world's most widely consumed psychoactive drug,
but unlike many other psychoactive substances, it is legal and unregulated in nearly all parts of
the world. Beverages containing caffeine, such as coffee, tea, soft drinks, and energy drinks,
enjoy great popularity. In North America, 90% of adults consume caffeine daily.
Part of the reason caffeine is classified by the Food and Drug Administration as GRAS
(generally recognized as safe) is that toxic doses (over 1 gram for an average adult) are much
higher than typically used doses (less than 500 milligrams). Ordinary consumption can have low
health risks, even when carried on for years there may be a modest protective effect against
some diseases, including Parkinsons Disease, and certain types of cancer. Caffeine can have both
positive and negative effects on anxiety disorders. Some people experience sleep disruption if
they consume caffeine, especially during the evening hours, but others show little disturbance
and the effect of caffeine on sleep is highly variable.
Caffeine increases cardiac arrhythmia (improper heart rate) by increasing stress hormone
(e.g.adrenaline) secretions. It has been shown there is an increase in brachial diastolic blood
pressure, but not in brachial systolic blood pressure. However, both aortic systolic and diastolic
blood pressures increase significantly during caffeine consumption. It has been noted that long
term consumption leads to increasing aortic systolic pressure which leads to chronic arterial
stiffness. The results of increasing blood pressure mostly contributes to blockage of Adenosine
A1 and A2 receptors. Since caffeine blocks adenosine A2 receptors which has vasodilatory

function, blood vessels become less dilated. (i.e. vasoconstriction) However, it is controversial
whether caffeine consumption increases heart rate. Some research shows that caffeine has no
influence on heart rate. It is hard to say how much dosage will cause increasing heart rate as no
studies have shown significant data. Different people have different tolerance for caffeine based
on individual metabolic activity, so there is no clear distinction between caffeine consumption
and the amount heart rate increases.

Problem statement:
How does caffeine affect the heart rate of Daphnia?
The heart rate of Daphnia sp. increases as the concentration of caffeine increases.
Manipulated : Concentration of caffeine
Responding : The heart rate of Daphnia per minute.
Fixed : Daphnia
Cavity slides, dropping pipettes, standard glassware (beakers, measuring cylinders etc) stop
watch, microscope.
Distilled water or pond water, 0.0%, 0.1%, 0.2%, 0.3%, 0.4% and 0.5% of caffeine solution,
Daphnia, cotton wool, paper towels.


1. Cavity slide is wiped with the paper towels to ensure the cavity slide clean from dust or
other microorganisms.
2. A few strands of cotton wool is placed on the cavity slide. This will
help restrict the movement of the Daphnia.
3. A large Daphnia is transferred from a beaker to a cavity slide by using a dropper.
4. Pond water is added onto the cavity slide by using a dropper to ensure the Daphnia still
alive. Excess water is removed from around the Daphnia by using filter paper.
5. The Daphnia is viewed by using a microscope under low power. Focus on its heart which
can be seen through its translucent body.
6. A stopwatch is used to record the number of heart beats for 30 seconds.
7. A pencil is tapped on a piece of paper. Then, the marks are counted up after 15 seconds.
8. Pond water is added necessarily to prevent the Daphnia die when absence of water.
9. The experiment is repeated for three times to get the average heart rate of the Daphnia.
10. Step 4 until 9 are repeated by replacing the pond water with 0.1%, 0.2% and 0.3% of
caffeine solution to drop onto the cavity slide. (0.1gm in 10ml = 0.1%)
11. For each concentration of caffeine, the experiment is repeated thrice to get the average
12. All the readings are tabulated in the provided table.


Concentration (%)
1st 15
2nd 15
3rd 15
4th 15
(heart beat
0 96 92 93 102 95.75 383
0.1 101 101 97 105 101 404
0.2 101 94 95 96 96.5 386
0.3 115 111 105 109 110 440
0.4 100 111 113 113 109.25 437
0.5 155 119 111 108 123.25 493

The experiment is executed to study the relationship between 6 different
concentrations of caffeine on the heart rate of Daphnia sp.. The result of this experiment is
recorded in the table above. The first experiment is taking the average heart beat of Daphnia sp
per 15 seconds. without the presence of caffeine and is set as the controlled experiment to

compare and contrast the result. The experiment is repeated for four times in order to get precise
value. The average result of the experiment without caffeine is 95.75 beats per 15 seconds. The
result then is multiplied by 4 to get the heart beat per minute which we got 383 heart beat. Next,
the experiment with caffeine presence in 0.1g/ml, 0.2g/ml, 0.3g/ml, 0.4 g/ml and 0.5g/ml result
with 404, 386, 440, 437 and 493 respectively.
In determining the effect of caffeine, we use Daphnia sp., an invertebrate, to carry out the
experiment. The ethical issue may arise here. However, instead of using a vertebrate, it is wiser
to use an invertebrate like Daphnia sp. This is because it has less nerves or simpler nervous
system and the pain is less throughout the experiment or may not feel pain at all. In addition,
Daphnia sp. is used as it is easier to see the differences or changes in its heartbeat because of its
translucent body.
There are some safety precautions that should be taken into account while carrying out
this experiment. First, the volume of water surrounding the Daphnia sp. on the cavity slide
should be reduced to avoid much movement of escape of Daphnia sp. Other than that, the light
of the compound microscope should be turned off when it is not used as live Daphnia sp. can be
killed at temperature of 40C. A few strand of muslin wool should be placed on the cavity slide
to prevent movement of Daphnia sp. making it easier to see under compound microscope. The
concentrations of caffeine solution must be used begin with low concentration by using 0.1%
concentration because Daphnia sp. is easily died if there is presence of high concentration of
caffeine to ensure that the Daphnia can adapt the changes in its culture.
There are some limitation during the experiment is carried out. The position of the heart
of the live Daphnia sp. cannot be located accurately. Therefore some students estimate the

heartbeat by counting the number of beats of legs or the contraction of the body. The
assumptions using the beats of the legs or the contraction of the body is the number of the upper
part of the thorax is closely to the heart. Another limitation is most of the students cannot used
the same Daphnia sp. to count the heart beat because Daphnia sp. is easily died as it is immersed
in excess concentration of caffeine. Hence the Daphnia sp. become weaken and eventually died.
The results obtained are reliable because the experiment is repeated for four times for
different conditions in different concentrations and the average reading of the heart beat are
counted. The reading also shows increasing in the heartbeat of daphnia sp. when there is the
presence of caffeine.
We should use a healthy Daphnia in the experiment to obtain a more accurate reading.
We should also try not to disturb the Daphnia sp. as much as possible as when we scared it, the
heart rate of the Daphnia sp. would not be accurate.

The heart rate of Daphnia sp. increases as the concentration of caffeine increases.


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