Ms Isaac

Kevin Pereira

The Effect Of Caffeine On The Heart Rate of

Daphnia.
Plants produce caffeine as an insecticide. Cocoa in South
America, coffee in Africa and tea in Asia have all been used for
hundreds of years to produce pick me up drinks containing
Caffeine. These days, caffeine is also used as a flavour enhancer
in a wide range of cola and other soft drinks. In addition, it has
medicinal uses in aspirin preparations, and is found in weightloss drugs and as a stimulant. In humans, caffeine acts as a
stimulant drug, causing increased amounts of stimulatory
neurotransmitters to be released. This also causes it to have an
effect on the heart. In this experiment we aim to see how
Caffeine concentration affects the heart rate of Daphnia.
Daphnia are a kind of small organism found in water, similar to
a prawn or insect, which has completely transparent skin,
making it easy to see the heart, and thus count the rate of its
heart beat. This is one of the main reasons we use Daphnia in
this experiment. It is also much easier and cheaper to obtain
results from them compared to doing the experiment on
humans for results, though they might not be as accurate. We
decided to carry out this experiment as a double blind test, to
make the results more accurate and less biased. We used two
solutions, one with normal coffee and the other with decaffeinated coffee. Both were put in unlabeled beakers, so that
nobody knew which solution was in which beaker. Normal
coffee would act as a stimulant drug and increase the heart
rate of the Daphnia, we assumed de-caffeinated coffee would
not affect the heart rate of the Daphnia since most of the
caffeine is removed, but after further research we realized that
de-caffeinated coffee is also a stimulant drug and would
increase the heart rate of the Daphnia as well.
Aim: To determine the effect of Caffeine on the heart rate of
Daphnia.
Hypothesis: The heart rate of the Daphnia will increase when it
is exposed to Caffeine and will keep increasing with exposure to
higher concentrations of Caffeine.
Substances and Equipment Needed:

Ms Isaac

Kevin Pereira

Culture Of Daphnia
Distilled Water
Caffeine Solution
De-caffeinated Coffee Solution
Paper Towels
Droppers
Light Microscope
Glass/Cavity Slides
Counter
Stopwatch
Calculator
50ml beaker
Measuring cylinder
Variables in this experiment:
Manipulated Variable: Presence of Caffeine
Dependent Variable: The heart rate of the Daphnia
Independent Variable: Surrounding temperature and light
intensity
The surrounding temperature and light intensity should be kept
constant because any changes could result in an inaccurate
heart rate, for example, high light intensity could stress the
Daphnia out and cause their heart rate to increase and give
inaccurate results.
Plan:
1. Take a small piece of cotton wool and keep reducing its
size until it is just a few fibres.
2. Place the fibres on a glass slide.
3. Add the Daphnia and some water until the Daphnia is just
covered by it.
4. Put the slide under a light microscope and adjust the
magnification so that the organs of the Daphnia can be
seen clearly.
5. Identify the heart of the Daphnia through the light
microscope.
6. Add a few drops of ethanol to slow down the heart rate of
the Daphnia if it is too fast to count.
7. Start a stopwatch and record the number of heartbeats
with a counter for 20 seconds.

Ms Isaac

Kevin Pereira

8. Repeat step 7 at least 3 times and record it in a table

along with the calculated mean.
9. Repeat steps 1-6 again with another Daphnia of similar
size to the one used previously.
10.
Add 5 drops of one of the unmarked solutions and
repeat steps 7 and 8.
11.
Repeat steps 1-6 again with a third Daphnia of similar
size.
12.
Add 5 drops of the second unmarked solution and
repeat steps 7 and 8
Risk Assessment: The test tubes and beakers should be
handled with care, to avoid damaging or breaking them and
harming the people using them and the people around them.
The light microscope should also be used with caution because
they are expensive pieces of equipment and are quite heavy,
they should be placed on proper tables so that they do not drop
on anybody and cause injuries. If precautions are taken to
ensure that these accidents dont happen, there should be no
incidents, because apart from these pieces of equipment there
isnt anything else that could cause harm. No hazardous
chemicals were used during this experiment so goggles and lab
coats did not need to be worn. The temperature and light
intensity should also be monitored carefully as there is a
chance for the daphnia to die when they are exposed to light
and heat which they arent used to. There were a few ethical
issues in this experiment as the Daphnia are living organisms
and died by the end of the experiment. This could raise some
ethical issues as they were killed for the experiment, but it does
not really make a big difference because they have short life
spans and are plentiful in nature.

Ms Isaac

Kevin Pereira

Ms Isaac

Kevin Pereira

Ms Isaac

Kevin Pereira

Analysis: By looking at the bar chart we can see that the mean
of the heart rate of the Daphnia with exposure to Caffeine
(assumed to be caffeinated solution) was only slightly more
than the heart rate without Caffeine(assumed to be
decaffeinated solution). The chart shows the mean of the
heartbeats of the Daphnia with exposure to the different
solutions, which is good because it shows the results more
clearly, in the table some values are much lower than others
and some values are higher in the control than in the caffeine
graph, which make the results seem inaccurate. The bar chart
is likely to look like this as decaffeinated also contains
stimulants in it and still contains a small amount of Caffeine,
this wouldve meant that the heart rates of the Daphnia would
still increase even without the Caffeine solution. This could also
be due to different sizes of Daphnia used and also Daphnia that
are of different ages. For example, younger Daphnia could have
faster heart rates than Daphnia that might be nearing the end
of their lifespan. The bar chart could also be slightly inaccurate
because alcohol was added at the start to decrease the heart
rate to make it easier to count. Different amounts of alcohol
given to the different Daphnia could have caused some heart
rates do decrease more than the others and thus show a lower
heart rate in the Caffeine graph. Another reason that the lines
might not be accurate is the amount of Caffeine added could
have varied slightly as it wasnt measure in definite amounts,
rather in drops and some drops could have been greater than
others and thus affect the heart rate.
Improvements:
1. Use another control solution as well that contains no
stimulants, such as water.
2. Use Daphnia that are all roughly the same size or the
same mass.
3. Add equal amounts of alcohol and solution (caffeinated or
decaffeinated) to each Daphnia slide.
Conclusion: The heart rate of the Daphnia does increase with
Caffeine as seen in the bar chart, even though it is just slightly
as Caffeine is a stimulant and affects the nervous system. I
have also concluded that de-caffeinated coffee does increase
the heart rate even though 97% of the caffeine concentration is

Ms Isaac

Kevin Pereira

reduced as caffeine is not the only stimulant in coffee and there

are other factors that affect the nervous system and increase
the heart rate in coffee as well.