Does caffeine affect heart rate?

Purpose: To investigate the effect of caffeine on the heart rate if Daphnia.

Introduction: Caffeine acts as a stimulant by exerting an effect on the central nervous
system. Caffeine stimulates receptors located in cells within the heart to increase the
heart rate. Effects of this stimulation speed up blood flow because of an increase in
heart rate.
Hypothesis: As caffeine is a stimulant, it will increase the heart rate of the daphnia,
as impulses become faster.
Apparatus:
-Concentrations of caffeine (0.1-0.5%)
-Microscope
-Culture of Daphnia
-Cavity slide
-Pipette
-Distilled water
-Cotton wool
-Stop clocks
Procedure:
1. Take a small piece of cotton wool and place it on a cavity slide.
2. Select a large Daphnia and use a pipette to transfer it onto the cotton wool fibers.
3. Add distilled water to the Petri dish until the animal is just covered by the water.
4. Place the Petri dish on the stage of a microscope and observe the animal under low
power. The beating heart is located on the dorsal side just above the gut and in front of
the brood pouch. Make sure that you are counting the heartbeats, and not the flapping
of the gills or movements of the gut. The heart must be observed with transmitted
light if it is to be properly visible. This is the control test (0% caffeine)
5. Use a stopwatch to time 30 seconds, and count the number of heartbeats in several
periods of 30 seconds. Count the beats by making dots on a piece of paper. Count the
dots and express heart rate as number of beats
per minute.
7. Add one drop of 0.1% caffeine to the cavity
slide with daphnia. Record the rate of heart beat
again.
8. Repeat this test using concentrations of 0.2%
to 0.5%
Results:
Concentration
(%)
0
0.1
0.2
0.3
0.4
0.5

Beats Per Minute (BPM)

Avera
3
1
2
ge
148 272 226 215
178 280 280 246
156 272 180 203
134 300 242 225
170 298 244 237
180 316 254 250

Graph:

Average Beats Per Minute (BPM)

300
250
200
BPM 150
100
50
0
0.0

0.1

0.2

0.3

0.4

0.5

Caffeine concentration (%)

Risk assessment:
Care should be taken when mixing solutions. All safety hazards and guidelines that
apply to any chemicals used should be followed. Make sure no Daphnia are killed in
the procedure of taking them out of the culture.
Evaluation:
We took into consideration that the Daphnia could adapt to the environment by using
different Daphnia for each experiment. Also, the amount of caffeine was maintained
constant to increase validity of the results (one drop). The measurements are made
less reliable due to the fact that they were repeated but not with great accuracy (the
results were very different each time). The heart rate of a Daphnia without caffeine
should be around 250 beats per minute, and when we measured them the mean value
was 215, therefore showing that our results are very quite from the norm.
Sources of potential error are:
-Human error-the subject timing will always react too late as time will have passed
when they read 30 seconds and when they say that 30 seconds have passed.
-The subject counting the number of heartbeats will have human error as they are
looking through the microscope and counting the amount of visible heartbeats. The
heartbeats are very fast, so it is very likely that many are missed. To prevent this, we
should have used a stroboscope so that readings would have been much more
accurate.
-Moreover we found that some Daphnias had a very slow heart rate when taken out
of the culture. This may be due to the fact that they were weak or dying. This will
affect the results, as the heart will need to work hard to keep the circulation going, so
the daphnia will have a higher risk of being affected.
-The time allowed for the daphnia to equilibrate will make a difference as well as they
may not have gotten used to their environment.
-We also didnt standardize what daphnia we used (sex/size). Usually, there is
individual variation so results will vary according to size(etc.)
-Human bias must be taken into account, as subjects will be expecting the heart rate to
increase, so have a tendency of making results support their hypothesis. Therefore

blind experiments must be done so that subjects dont know what they are testing
(i.e. 0.1% caffeine or no caffeine).
Ethical issues:
Organism has no consent and the nervous system may be harmed. We are not able to
prove that they do not feel anything. Also the value of the study itself has to be taken
into account. If thousands of students do this experiment, many daphnia will die.
These ethical issues will be taken into account by returning the Daphnia to the beaker
after being used, storing the daphnia in conditions that replicate its natural
environment. Also, Daphnia have not got a developed nervous system, so do not feel
the same sort of pain and they are easily available (they can be bred and are not in risk
of becoming extinct.)
Dependent and independent variables:
The heart rate of the daphnia is the dependent variable, and the concentration of the
caffeine is the independent variable. Controlled variables are the amount of solutions
added (in this case 1 drop from a pipette) to increase accuracy.
Conclusion:
There is an overall increase of the heart rate of Daphnia as the concentration
increases, for example at 0% the heart rate is 215, whereas with 0.3%, the average is
225. So we can see that the trend is a positive increase (positive gradient). But we can
also see that there is one anomalous result (0.2%) where there is a dip in the graph.
This occurs in all three repetitions of the experiments. This could be due to bias in
results, as one subject had this; the others expected this to happen as well, or simply
due to human error, or due to having a weaker daphnia in each case. This is why it is
more reliable to do many repetitions, making the experiment more repeatable.
The hypothesis suggested has been verified, although the experimental method was
not very valid. Therefore no real valid conclusion can be drawn from the results.
Background knowledge-Caffeine:
Caffeine mimics some of the effects of adrenaline (and noradrenaline) in the heart. It
increases the amount of cAMP in the sinoatrial node. As cAMP levels increase, the
electrical activity of the sinoatrial node increases, making it depolarize and 'beat'
faster. Caffeine has additional effects on the heart. Like adrenaline, it can affect the
main pumping chambers (ventricles), leading to an increase in the rate of contraction
and relaxation of each heartbeat. This means that, as well as beating faster, the heart's
individual beats are associated with an increased volume of blood ejected into the
circulation per unit time. This is called increasing cardiac output. Two or three cups of
strong coffee or tea contain enough caffeine to cause an increase in human heart rate
of 5-20 beats/min.