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AIM : To investigate the effect of caffeine on heart rate HYPOTHESIS : There is relationship between the concentration of caffeine and heart rate of Daphnia. When the concentration of caffeine increases, the heart rate of Daphnia will increase as well. INTRODUCTION : This experiment deals with the affects of caffeine on the heart rate of daphnia. The following includes information about daphnia and caffeine. Caffeine medically known as trimethylzanthine. Its chemical formula is C8H10N4O2. When in pure form, caffeine is a white crystalline powder that taste very bitter. The most common way of acquiring pure caffeine is the process of decaffeinating coffee and tea. Caffeine is useful as a cardiac stimulant and also as a mild diuretic. Cardiac stimulants increase the heart rate, and diuretics increase urine production. Caffeine is also used to provide a boost of energy or a feeling of heightened alertness. Caffeine is an addictive drug. It operates using the same mechanisms that amphetamines, cocaine and heroin use to stimulate the brain. Caffeines effects are more mild than the above listed drugs, but it is manipulating the same channels and this gives caffeine its addictive qualities. If you feel like you cannot function without it, then you are addicted to caffeine. Caffeine occurs naturally in many plants, such as, coffee beans, tea leaves and cocoa nuts. Caffeine is added artificially to many products, including a variety of beverages. Your brain creates a chemical called adenosine. This chemical binds to adenosine receptors in the brain. The binding of adenosine causes drowsiness by slowing down nerve cell activity. To a nerve cell, caffeine looks

like adenosine, and caffeine binds to the adenosine receptors. The caffeine does not slow down the cells activities like adenosine, but instead, speeds up the cells activities. Caffeine also constricts the brains blood vessels. If you have a vascular headache, caffeine can slow down the blood flow to the brain and relieve it. Therefore, some headache medicines like Anacine contain caffeine. The pituitary gland sees all of the activity and thinks some sort of emergency must be occurring and causes a release of the hormone adrenaline. Adrenaline speeds up the bodies functions even more. Figure below show the structure of caffeine. The most important long-term problem is the effect that caffeine has on sleep. Adenosine reception is important to sleep, and especially to deep sleep. If you drink a big cup of coffee with 200 mg of caffeine in it at 3:00 PM, about 100mg is still in your system at 9:00 PM. You may be able to fall asleep, but your body probably will miss out on the benefits of deep sleep. The deficit adds up fast. The next day you feel worse, so you need caffeine as soon as you get out of bed. The cycle continues day after day.

IUPAC name

1,3,7-trimethyl- 1Hpurine- 2,6(3H,7H)dione 1,3,7-trimethylxanthine, trimethylxanthine, theine, methyltheobromine

Other names

Daphnia, also commonly called water fleas are small aquatic crustaceans. They are about 0.3 mm. They are freshwater zooplankton and consume phytoplankton. Some daphnia are, however, carnivorous, eating other zooplankton. The body of a daphnia are laterally compressed. That is, they are flat. The division of the body into segments is nearly invisible. The head is fused, and is generally bent down towards the body with a visible notch separating the two. In most species the rest of the body is covered by a carapace, with a ventral gap in which the five or six pairs of legs lie. The most prominent features are the compound eyes, the second antennae, and a pair of abdominal setae. In many species, the carapace is translucent or nearly so and as a result they make excellent subjects for the microscope as one can observe the beating heart. Their bodies are divided into three parts, similar to an insect. The part I will be concentrating on is the heart. The heart of a daphnia is rather conspicuous .It is easily seen while viewing the crustacean under a microscope. Because the heart is so large and its beating can be easily counted, daphnia are commonly used in a variety of physiological experiments including response to temperature changes and responses to stimulants and depressants.

The heart rate of human may change depends on the bodies condition. Like human, the heart rate of Daphnia may change due to the differences of caffeine concentration. Too low caffeine concentration would not give effects to Daphnia but too high caffeine concentration could be fatal to them. Therefore, a pilot experiment can be done to establish the concentration range that will be testing in the full experiment. In other words, at the lowest concentration we see no response, and at the highest concentrations we see a maximal response, with several intermediate responses in between. The pilot experiment will help to determine the minimum and maximum concentrations to use. Then, the heart rate of Daphnia can be counted by placing it in the known concentration of caffeine and put it below a light microscope. ICam also can be used to help the counting work, but here I used to count myself as ICam is not provided in the lab.

MATERIALS AND APPARATUS : Culture of Daphnia, distilled water or pond water, 0.1%,0.2%,0.3%,0.4% and 0.5% caffeine solution, cavity slide, dropping pipettes, muslin cloth, beakers, measuring cylinder, stopwatch, microscope, spatula.

PROCEDURE : 1. 0.1%, 0.2%, 0.3%, 0.4% and 0.5% caffeine solution are prepared. 2. A Daphnia is placed on the cavity slide on the microscope using a spatula. 3. The position of the Daphnia is altered so that we can observe it from the side view using muslin cloth. 4. A drop of pond water is dropped onto the Daphnia using a dropping pipette.

5. The heart rate of Daphnia is counted twice and the average heart rate is counted. 6. The experiment is repeated using 0.1%, 0.2%, 0.3%, 0.4%, and 0.5% caffeine solution instead of using pond water. 7. All data of the results is recorded. 8. A graph of heartbeat of daphnia per minute against concentration of caffeine is plotted.

RESULTS : CONCENTRATION OF CAFFEINE SOLUTION (%) 1 2 3 AVERAGE STANDARD DEVIATION 0.0 0.1 0.2 0.3 0.4 0.5 8 17 24 32 36 40 8 19 15 25 33 36 15 14 15 11 33 32 10.30 16.67 18.00 22.67 34.67 36.00 3.40 2.03 4.24 8.72 6.60 3.27 HEART RATE OF DAPHNIA MAGNA WITHIN 20 SECONDS

CONCENTRATION OF CAFFEINE SOLUTION (%) 0.0 0.1 0.2 0.3 0.4 0.5


30.90 50.01 54.00 68.01 104.01 108.00

The graph of heart rate of Daphnia per 20 seconds against caffeine concentration
40 35 30 25 20 15 10 5 0 0 0.1 0.2 0.3 0.4 0.5 Heart rate of Daphnia per 20 seconds


120 100 80 60 Heart rate of Daphnia/minute 40 20 0 0 0.1 0.2 0.3 0.4 0.5 Concentration of caffeine (%)

ANALYSIS OF DATA : 1. Standard deviation of the data is calculated using the formula x2 - x2 n where x=1,2 and 3 reading n=number of reading x2=square of average value 2. Both graph of heart rate of Daphnia against caffeine concentration show that the heart rate of Daphnia increases as the caffeine concentration increases.

DISCUSSION : From the graph of heart rate of Daphnia per minutes against caffeine concentration, the heart rate of Daphnia increases generally as the caffeine concentration increases. This is because just like human, a daphnias heart will beat faster when it receives a dose of caffeine. Caffeine belongs to a class of compounds called methylxanthines and can block a receptor on the surface of heart muscle cells for adenosine. In fact, it is caffeine's blockade of the A1 adenosine receptor in the heart that causes the heart to pound after a significant caffeine dose. Caffeine and similar compounds also inhibit a class of enzymes known as cyclic nucleotide phosphodiesterases. These enzymes are, in part responsible for degrading a stimulatory signal produced when excitatory neurotransmitters activate different neurons in the central nervous system (CNS). Thus, when they are inhibited by caffeine, the stimulatory signal remains active for a longer period of time resulting in a greater sense of alertness (a CNS effect) but also a higher heart rate, blood pressure and respiratory rate. Therefore, the heart rate of Daphnia increases.

In this experiment, concentration of caffeine acts as an independent variable. It is manipulated throughout the experiment by using different volume of caffeine in a solution to get different heart rate of Daphnia, which is the dependent variable. The other factors including time taken, pond water, culture of Daphnia and temperature are kept constant. Under the condition of all the other variables kept constant as possible, the heart rate become higher due to the increase of caffeine concentration, since more caffeine molecule available to react with the body. A control experiment is carried out using 0% caffeine concentration as a comparison. This is to prove that any reaction that happen is due to a specific factor. An experiment without caffeine is to show the normal heart rate of Daphnia. However, there are some errors occur in data. This may be due to some experimental error or rather limitation of the experiment. The caffeine concentration may not be accurate. This is because the different concentration of caffeine may mix together while we test the Daphnia. For example, 0.1% caffeine concentration is dropped onto the Daphnia. Then, a muslin cloth is used to dry the Daphnia so that we can proceed the experiment using the other caffeine concentration, but there will got some of the 0.1% caffeine concentration remains at the body of the Daphnia because it is not washed or fully cleaned to remove the previous caffeine concentration. Therefore, the 0.2% caffeine concentration will mix with the remaining 0.1% caffeine concentration that are not wash out. This will increase the percentage of caffeine in the 0.2% caffeine concentration. Besides that, the temperature of the surrounding also affects the Daphnia. When the Daphnia is put on the microscope for a long time, the temperature of the surrounding will increase due to the presence of light at the microscope. Here, the Daphnia would react due to the different caffeine concentration and the surrounding temperature.

In order to obtain a more accurate result, the errors must be controlled. First, used muslin cloth instead of using spatula to coordinate the Daphnia. If we use spatula, the Daphnia may injured because it is a very small animal and can be easily injured. We also should prevent using tap water to keep the Daphnia because tap water usually contains chlorine as a disinfectant which will kill Daphnia and other aquatic organism. The chlorine may change the condition of the Daphnia before the experiment. Other than that, after testing the Daphnia with one caffeine concentration, let the Daphnia swim in the pond water for a minute to wash out any caffeine left on its body and to maintain the Daphnias temperature. Using this way, we can have a fresh Daphnia for the second experiment using the next caffeine concentration. Hence, we can get a more accurate heart rate.

CONCLUSION : The heart rate of Daphnia increases as the concentration of caffeine solution increases provided that the concentration is not too high which could be fatal to Daphnia. The hypothesis is accepted. REFERENCES : 1. Edexcel Biology For AS, Hodder Education, C.J Clegg, 2008. 2. Edexcel AS Biology, Pearson Education Limited, 2008. 3. http://en.wikipedia.org/wiki/Main_Page 4. http://www.madsci.org/posts/archives/1999-07/931925101.Zo.q.html 5. http://www.sciencebuddies.org/science-fair-projects/project_ideas.shtml