UNIT 1

1. The Effect of Caffeine on Heart Rate

Why Daphnia?

- Daphnia are small and show the results of the experiment quickly
- They have simple nervous systems so are less likely to feel pain
- They are abundant so easy to get hold of / No damage to environment when a few are
removed from it
- Transparent so easy to measure heartbeat

· Variables to be controlled are:

- Temperature
- Volume of solution
- Time of acclimatisation
- Daphnia must be the same age / size
- Daphnia must have the same health
- Counting time

· Method:

- Independent variable: Caffeine concentration

- Dependent variable: Heart rate of Daphnia
- Place a Daphnia in each of 5 different solutions (4 different concentrations of caffeine and
one with distilled water to act as a control)
- Leave Daphnia for 5 minutes to acclimatise
- Immobilize the Daphnia using a vaseline
- Count and record the no. of heartbeats in one minute
- Repeat 5 times at each concentration and allow means to be calculated
· Outcome: As caffeine concentration increases, heart rate also increases

· How does caffeine increase heart rate?

Increasing caffeine concentration causes the electrical activity of the sinoatrial node to
increase, making it depolarize. As it depolarizes, the right and left atria contract and the
impulse travels to the atrioventricular node where, after a delay of about 0.13 seconds, the
impulse continues to travel towards the ventricles. This delay ensures that the atria have
finished contracting and ventricles are full. The signal then reaches the Purkyne fibres that
conduct the impulses to the apex of the ventricles where contraction begins and travels
upwards towards the atria.

Caffeine also affects the ventricles, leading to an increase in the rate of contraction and
relaxation of each heart beat. This means that, as well as beating faster, the heart's individual
beats are associated with an increased cardiac output.
2. The vitamin C Content of Fruit Juice

· Variables to be controlled:

- Mass of fruit / age / source / …

- Time for storage
- Method of juice extraction
- Volume / concentration of juice / DCPIP
- Temperature
- Same end point colour

· Method:

- Independent variable: Fruit juice

- Dependent variable: Volume of juice required to decolourise 1cm3 of DCPIP
- Put 1cm3 of DCPIP solution into a test tube
- Fill a plastic syringe with juice and add drops to the DCPIP until the blue of the DCPIP is
lost. Record the volume of juice added
- Repeat 5 times for each juice to calculate means
- To calculate the actual Vitamin C concentration, the DCPIP solution must be calibrated. A
solution of known Vitamin C concentration is added to 1cm3 of DCPIP until it is decolourised
and the volume recorded.
- Conc. of Vitamin C in juice = (Con. of Vitamin C solution x Volume of Vitamin C solution
needed to decolourise 1cm3 DCPIP) ÷ Volume of fruit juice needed to decolourise 1cm3 DCPIP

· Limitations:
- Difficulty in controlling temperature

- End point difficult to judge as needs to be just when blue colour disappears especially in
highly coloured juice
- Some loss of solution when transferring from one beaker to another

- Accuracy of measuring equipment

· Storing fruit at low temperatures slows down decay because:

- Low temperature reduces / prevents growth of microorganisms
- Low temperature reduces activity of enzymes
- Less kinetic energy means fewer collisions / fewer cell divisions
3. The effect of Temperature on Membranes permeability

· Factors that affect the permeability of the beetroot cell membrane are:

- Temperature
- Age
- Storage
- Duration
- Pre treatment with solvent
- pH

· Variables to be controlled are

:
- Source / Species of beetroot
- Age / Size of beetroot
- Volume of water / solution used
- Time left in water or solution
- pH 


· Method:

- Independent variable: Temperature of water


- Dependent variable: % transmission of light through resulting solution
- Using a cork borer and knife, cut 5 pieces of beetroot equal in mass and size
- Rinse the beetroot pieces with water and gently pat them dry with tissue before using
them as, when cutting, pigment is released from broken cells and must be removed before
starting or solutions will be darker than they should
- Place one piece into each of 5 tubes and add 5 1cm3 water to each one
- Place each tube into a water bath of different temperature (e.g. 15, 20, 25, 30, 35)
- Leave for 15 minutes
- Remove beetroot and shake tubes to disperse dye.
- Calibrate the colorimeter using distilled water in a cuvette as a reference / control.
- Take readings of absorbance of the water in the tubes
- Take 5 repeats at each temperature and calculate means

· Results and reasons:

As temperature increases, % transmission slightly increases. This is due to membrane molecules

gaining more heat energy and vibrating more, creating large gaps in the membrane that enable
dye to be released. Proteins in membrane may become denatured, leaving large pores through
which the dye leaks.

· Limitations:

- Pigment is not equally distributed throughout the beetroot

- Some beetroot may have skin on affecting surface area
- Size of beetroot is difficult to control
4. The Effect of Enzyme Concentration on the Rate of Reaction

· Variables to be controlled are:

- Temperature à use a water bath

- Volume of enzyme
- Volume of substrate
- Concentration of substrate

- pH

· Method:

- Take 5 test tubes. In 4, place increasing volumes of trypsin solution e.g. (1, 2, 3, 4 cm3 ) and
make up the volume to 4 cm3 using distilled water. The other test tube should be filled with
4 cm3 of water to act as a control.
- Add 5 of milk powder (casein solution) as substrate and start the stopwatch
- Measure the cloudiness of the solution over time using a colorimeter (every 30 secs for 10
minutes) against water as a reference / control
- Repeat at least 5 times at each concentration and calculate means
· As concentration of enzyme increases, rate of reaction increases up to a point, where all
enzyme has metabolised all substrate immediately

· Why do enzymes work better at higher temperatures?

At low temperatures the reaction is slow because the enzyme and substrate molecules don’t
collide very often and move slowly. Increasing temperature increases kinetic energy and so
frequency of collisions. The substrate binds to the enzyme’s active site more often thus
increasing the rate of reaction. After the optimum temperature bonds holding the 3D – shape
of the enzyme together start breaking so it loses its shape and the enzyme-substrate complex
can no longer form. The enzyme is denatured.