Aelfgar College

Biology AS

Investigating the effect of

caffeine on heart rate
Anna Phizacklea
Introduction
Aim: to use Daphnia to determine whether caffeine concentration affects on heart rate.

Hypothesis: As the caffeine concentration increases, the heart rates of the Daphnia will
also increase.

Independent variable: Caffeine concentrations

Dependent variable: The heart rate of the Daphnia

Control variable Possible effect Control measure

Knowledge of the caffeine If the person counting the
concentration heartbeats knows what the
concentration of caffeine is,
they may be inclined to count
more or less heartbeats
subconsciously, due to their
expectations.
Water temperature At colder temperatures,
Daphnia move less and have a kept constant at around room
lower heart rate to conserve
energy. At higher temperatures,
they will move about more and
have a higher heart rate as
they have excess energy.
Light exposure If kept under the microscope
light for too long, they may
begin to overheat and die. This
means the daphnia may not be
available for later trials.
Length of exposure to caffeine If one daphnia has longer
exposure to the caffeine
solution than another, it may
have a greater effect, even if
the caffeine solution is of a
lower concentration. Too much
caffeine may affect its health.
Daphnia used There are genetic differences
between each daphnia, so
caffeine may affect each one
slightly differently.
A blind trial – the person
measuring out the caffeine
solutions should not tell the
person counting the heart rate
what concentration it is.
Water temperature should be
temperature.
The length of exposure should
be minimised – when not
counting the heartbeats, the
light should be turned off.
Organise timing between the
group and stick to it, e.g. after
30 seconds wait, heartbeats
should be counted over 10
seconds, wait for 20, and
repeat 3 times.
Ideally, the same daphnia
should be used for each trial.
However, this is unrealistic:
there may be contamination
from previous caffeine
concentrations, leaving lasting
results; the daphnia may die
from too much exposure to
caffeine and light.

Ethical issues: There is a problem with this particular experiment of whether it is right to
use living organisms. At the time there is not a better alternative, so steps will be taken to
minimise suffering to the Daphnia, i.e. their exposure to bright lights, heat and caffeine will
be minimised as much as possible.
Background information: 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 weight-loss drugs and as a stimulant in
students' exam-time favourites like Pro-plus and Red Bull.
In humans, caffeine acts as a stimulant drug, causing increased amounts of stimulatory
neurotransmitters to be released. At high levels of consumption, caffeine has been linked
to restlessness, insomnia and anxiety, causing raised stress and blood pressure. This can
lead to heart and circulation problems.

Equipment and materials

 Culture of Daphnia (water fleas)
 Cavity slides
 Dropping pipettes
 Distilled water
 Caffeine tablets
 Cotton wool
 Standard glassware (beakers, measuring cylinders)
 Stop-clock
 Paper towels or filter paper
 Microscope

Method
 A variety of caffeine concentrations should be made using caffeine tablets and
water. For this example, 0.1%, 0.2%, 0.3%, 0.4% and 0.5%. 0% (water) should also
be used as a control.
 Using a dropping pipette, a small volume of the first caffeine solution should be
placed onto a cavity slide (e.g. 5 drops) by one member of the pair – the person
counting should be unaware which solution is being used.
 Using another dropping pipette, a water flea should be picked up from the culture.
Squeeze gently to release as much excess water as possible, before dabbing away
the remaining using cotton wool or paper towels.
 Place the water flea into the caffeine solution on the cavity slide and start the stop-
clock. Place under the microscope.
 Leave for 30 seconds to allow the solution to take effect.
 Count the heartbeats in 10 seconds under the microscope. Turn the light off to
minimise damage to the daphnia, wait 20 seconds and repeat as necessary to get a
sufficient number of results (e.g. 4).
 Repeat using different caffeine solutions.
Results

Raw data
Heart beats counted in 10 seconds
Concentration of 1 2 3 4 Mean
caffeine solution
0% 31 28 33 36 32
0.1% 46 45 42 44 44.25
0.2% 51 50 46 48 48.75
0.3% 48 47 52 49 49
0.4% 52 51 49 51 50.75
0.5% 51 55 50 53 52.25

Conclusion
It is clear from the results that as the concentration of the solution increases, the heart rate
of the organism also increases. There is a significant increase in heart rate from 0% to
0.1%, though the heart rate increases at a slower rate between the concentrations. This
suggests there is a limit to how much caffeine can be taken in by the Daphnia, so caffeine
affects the heart rate up to a certain point (higher concentrations needed to find when).

Evaluation
There were numerous flaws and sources of error throughout the experiment. A large flaw
in applying this to humans is that the Daphnia used are nowhere near humans in
evolutionary terms – the species are not remotely related. This means that for the general
assumption, that caffeine increases heart rate, to be reliable, the experiment would have to
be conducted again with humans and other species with an altered method (e.g.
consuming caffeine and measuring pulse). However, this produces more ethical issues, as
animals closer to humans genetically are generally more intelligent.
One of the biggest sources of error was the counting of the heartbeats. The main problem
with this was that the Daphnia moved a lot, so it was difficult to count steadily. In some
cases, students had to wait until they were still, which meant their exposure to caffeine and
light increased, affecting the results. Another problem was that the heartbeat was so quick,
it was either difficult to keep track of the heartbeat, or there may have been discrepancies
between groups of when the heart was beating, leading to differences in results. A final
problem with counting the heartbeats was locating the heart – some groups had difficulty
finding the heart, which again resulted in prolonged exposure to the caffeine and light.
Another source of error, as mentioned in the control variables, is the genetics of each
individual Daphnia. Whilst ideally the same Daphnia should've been used, this is
impractical. This meant that there were genetic differences between the Daphnia used
each time, so some may have had higher tolerance to the caffeine. Similarly, some
Daphnia may have been more aware of what was happening and of themselves being
moved and kept in a small volume of liquid. This may have increased their heart rates. A
final source of error was the age/state of the Daphnia: for instance, whether the Daphnia
was pregnant, old, new-born, male or female would've had an effect on its heart rate
anyway, so the results wouldn't have been fully reliable. Again, ideally the same Daphnia
should've been used, or at least the same type of Daphnia – however, this would be very
difficult to determine.