Required Practicals 7 - 12 Flashcards
RP7 - What is the purpose of chromatography?
To separate different components in a sample.
RP7 - State the factors affecting the rate of migration of different pigments.
Solubility
Mass
Affinity to the paper
RP7 - What is the formula of the RF value?
Distance moved by pigment / Distance moved by solvent
RP7 - What is the purpose of finding the RF value of a pigment?
Experimental RF value can be compared to a standard value in a database to identify the pigment.
The standard value should be measured using the same paper and solvent.
RP7 - Outline the procedure of using chromatography to separate photosynthetic pigments.
- Draw a horizontal pencil line 1cm above the bottom of the filter paper.
- Add some acetone and use the mortar and pestle to grind up the leaf sample
and release the pigments. - Use a capillary tube to transfer the pigment onto the pencil line.
- Suspend the paper in the solvent so that the level of the liquid does not lie
above the pencil line and leave the paper until the solvent has run up the
paper to near the top. - Remove the paper from the solvent and draw a pencil line marking where the
solvent moved up to. - Calculate the Rf value for each spot.
RP7 - State the hazards and precautions in this practical.
Solvents are irritant and flammable.
Keep away from naked flames, wear eye protection and avoid contact with skin.
Leaf extract may be a biohazard.
Wash hands after use.
RP8 - What is the function of dehydrogenase in chloroplasts?
It catalyses the acceptance of electrons by NADP in the light dependent reactions.
RP8 - What is the purpose of DCPIP?
It is a redox indicator dye and acts as an alternate electron acceptor instead of NADP. It turns from blue to colourless when reduced.
RP8 - Why is the plant extract chilled in an ice-water bath?
To lower the activity of enzymes to prevent them from breaking down the
chloroplasts.
RP8 - How is the control set up?
Fill a cuvette with chloroplast extract and distilled water.
RP8 - How is light intensity controlled?
Adjust the distance of the lamp from the set up.
Perform the practical in a dark room so that the only light source is the lamp.
RP8 - What is the function of the muslin cloth?
To filter out any debris in the ground leaf mixture but allowing chloroplasts to pass through.
RP8 - Why are the stalks of leaves removed before grinding?
The stalks do not contain many chloroplasts.
RP8 - Outline the procedure of investigating the effect oflight intensity, after chloroplast extract has been obtained.
- Set the colorimeter to the red filter. Zero using a cuvette containing
chloroplast extract and distilled water. - Place test tube in the rack 30cm from light source and add DCPIP.
Immediately take a sample and add to cuvette. Measure the absorbance of
the sample. - Take a sample and measure its absorbance every 2 minutes for 10 minutes.
- Repeat for different distances from lamp up to 100 cm.
RP9 - What is the function of methylene blue in this practical?
It is a redox dye and acts as an alternate electron acceptor of the electrons transferred during ATP synthesis.
It turns from blue to colourless, indicating the end point.
RP9 - Outline the procedure to investigate the effect of temperature on the rate of respiration of yeast.
- Set up a water bath at 35°C.
- Add equal volumes of the yeast and glucose solution to three test tubes.
Place test tubes in the water bath and leave them to equilibrate for 10 minutes. - Add 2 cm3
of methylene blue to the test tubes and start the timer. Shake for 10 seconds and place test tube back in water bath. Record how long it takes for the methylene blue to turn colourless for each test tube. - Repeat the experiment using temperatures of 40°C, 50°C, 60°C and 70°C.