Photosynthesis Experiments

Using Chromatography to Investigate Photosynthesis

Chromatography is a technique that can be used to identify which pigments are in the leaves of different plants. This allows us to identify what wavelengths of light a plant can absorb.

1) Extract the chlorophyll

Take some leaves from a plant that grows in the shade.
Place boiling water from a kettle into a beaker and dip a few leaves to kill them.
Tear up the leaves and grind with 5-10 cm³ of acetone in a pestle and mortar until you obtain a dark green chlorophyll extract. Add a pinch of sand to break up the cells.

2) Add the chlorophyll

Use a strip of chromatography paper, which fits into a boiling tube without touching the sides. Mark a line 1.5 cm from the bottom in pencil and in the centre put a cross.
Mark another line 2 cm from the top of the paper.
Pick up your chlorophyll solution using a capillary tube and place on the cross. Allow to dry.
Repeat 4-5 times to have a small, dense green spot.

3) Suspend the paper

Put 1.5 cm of acetone/petroleum solvent into the bottom of the boiling tube, using a funnel to avoid wetting the sides.
Suspend the paper using a drawing pin so that it just touches the solvent and leave.

4) Repeat the experiment

Repeat steps 1-3 using leaves from a plant that grows well in direct sunlight.
Remove the paper from each boiling tube when the solvent reaches the line at the top of the papers. Mark the positions of the pigments before they fade.

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5) Calculate R_f

Calculate the R_f values using:
- R_f = distance travelled by spot ÷ distance travelled by solvent
Find the pigment that corresponds with the given R_f value.

Investigating Dehydrogenase Activity in Chloroplasts

During the light-dependent reaction, NADP is reduced to NADPH. A dehydrogenase enzyme catalyses this reaction. This experiment will monitor dehydrogenase activity by using DCPIP, a redox indicator.

DCPIP

In this investigation, a blue dye (DCPIP) is used to monitor the rate of dehydrogenase activity. DCPIP is a redox indicator.
This means that it is blue in the oxidised state and colourless in its reduced state.
When electrons are released by the chlorophyll, DCPIP will change from blue to colourless.

1) Extracting chloroplasts

Put 50 cm³ of isolation medium into a beaker.
Tear eight spinach leaves into small pieces and put the pieces into the isolation medium in the beaker.
- Do NOT put pieces of the midrib or the leaf stalk into the beaker.
Half fill a large beaker with ice and place a small beaker on top of the ice.
When carrying out this step, all solutions and apparatus should be kept as cold as possible and the extraction should be carried out as quickly as possible.

2) Suspending chloroplasts

Put 3 layers of muslin over the top of the filter funnel and wet with the isolation medium. Rest the filter funnel in the small beaker on the ice.
Pour the spinach and isolation medium into the blender and blend for 15 seconds. Pour the blended mixture back into the beaker.
Pour your blended mixture through the muslin in the filter funnel. Carefully squeeze the muslin to assist the filtering process.
Label this filtrate which is in the small beaker on ice as ‘chloroplast suspension’.

3) Set up tubes A and B

Label five test tubes A, B, C, X and Y and stand them in the large beaker. Put the lamp about 10 cm away so that all tubes are illuminated. Set up tubes A and B as follows:
- Tube A - 5 cm³ DCPIP solution + 1 cm³ water + 1 cm³ chloroplast suspension. Immediately wrap the tube in aluminium foil to exclude light.
- Tube B - 5 cm³ DCPIP solution + 1 cm³ water + 1 cm³ isolation medium.
Tubes A and B are control experiments. Leave both tubes until the end of your investigation.

4) Set up tube C

Set up tube C as follows:
- Tube C - 6 cm³ water + 1 cm³ chloroplast suspension.
Tube C is for you to use as a standard to help you to determine when any colour change is complete.

5) Set up tube X

Set up tube X as follows:
- Tube X - 5 cm³ DCPIP solution + 1 cm³ water in the tube.
Add 1 cm³ chloroplast suspension to tube X, quickly mix the contents and start the timer.
Record in seconds how long it takes for the contents of tube X to change colour from blue-green to green. Use tube C to help you determine when the colour change is complete.
- Repeat this step four more times.

6) Set up tube Y

Set up tube Y as follows:
- Tube Y - 5 cm³ DCPIP solution + 1 cm³ ammonium hydroxide.
Add 1 cm³ chloroplast suspension to tube Y, quickly mix the contents and start the timer.
Record in seconds how long it takes for the contents of tube Y to change colour from blue-green to green. Use tube C to help you determine when the colour change is complete.
- Repeat this step four more times.

7) Record the results

Record your data in a suitable table.
At the end of your investigation, record the colour of the mixtures in tubes A and B.

1Biological Molecules

1.1Monomers & Polymers

1.1.1Monomers & Polymers

1.1.2Condensation & Hydrolysis Reactions

1.2Carbohydrates

1.2.1Structure of Carbohydrates

1.2.2Types of Polysaccharides

1.2.3End of Topic Test - Monomers, Polymers and Carbs

1.2.4Exam-Style Question - Carbohydrates

1.2.5A-A* (AO3/4) - Carbohydrates

1.3Lipids

1.3.1Triglycerides & Phospholipids

1.3.2Types of Fatty Acids

1.3.3Testing for Lipids

1.3.4Exam-Style Question - Fats

1.3.5A-A* (AO3/4) - Lipids

1.4Proteins

1.4.1The Peptide Chain

1.4.2Investigating Proteins

1.4.3Primary & Secondary Protein Structure

1.4.4Tertiary & Quaternary Protein Structure

1.4.5Enzymes

1.4.6Factors Affecting Enzyme Activity

1.4.7Enzyme-Controlled Reactions

1.4.8End of Topic Test - Lipids & Proteins

1.4.9A-A* (AO3/4) - Enzymes

1.4.10A-A* (AO3/4) - Proteins

1.5Nucleic Acids

1.5.1DNA & RNA

1.5.2Polynucleotides

1.5.3DNA Replication

1.5.4Exam-Style Question - Nucleic Acids

1.5.5A-A* (AO3/4) - Nucleic Acids

1.6ATP

1.6.1Structure of ATP

1.6.2End of Topic Test - Nucleic Acids & ATP

1.7Water

1.7.1Structure & Function of Water

1.7.2A-A* (AO3/4) - Water

1.8Inorganic Ions

1.8.1Inorganic Ions

1.8.2End of Topic Test - Water & Inorganic Ions

2Cells

2.1Cell Structure

2.1.1Introduction to Cells

2.1.2Eukaryotic Cells & Organelles

2.1.3Eukaryotic Cells & Organelles 2

2.1.4Prokaryotes

2.1.5A-A* (AO3/4) - Organelles

2.1.6Methods of Studying Cells

2.1.7Microscopes

2.1.8End of Topic Test - Cell Structure

2.1.9Exam-Style Question - Cells

2.1.10A-A* (AO3/4) - Cells

2.2Mitosis & Cancer

2.2.1Mitosis

2.2.2Investigating Mitosis

2.2.3Cancer

2.2.4A-A* (AO3/4) - The Cell Cycle

2.3Transport Across Cell Membrane

2.3.1Cell Membrane Structure

2.3.2A-A* (AO3/4) - Membrane Structure

2.3.3Diffusion

2.3.4Osmosis

2.3.5Active Transport

2.3.6End of Topic Test - Mitosis, Cancer & Transport

2.3.7Exam-Style Question - Membranes

2.3.8A-A* (AO3/4) - Membranes & Transport

2.3.9A-A*- Mitosis, Cancer & Transport

2.4Cell Recognition & the Immune System

2.4.1Immune System

2.4.2The Immune Response

2.4.3Antibodies

2.4.4Primary & Secondary Response

2.4.5Vaccines

2.4.6HIV

2.4.7Ethical Issues

2.4.8End of Topic Test - Immune System

2.4.9Exam-Style Question - Immune System

2.4.10A-A* (AO3/4) - Immune System

3Substance Exchange

3.1Surface Area to Volume Ratio

3.1.1Size & Surface Area

3.1.2A-A* (AO3/4) - Cell Size

3.2Gas Exchange

3.2.1Single-Celled Organisms

3.2.2Multicellular Organisms

3.2.3Control of Water Loss

3.2.4Human Gas Exchange

3.2.5Ventilation

3.2.6Dissection

3.2.7Measuring Gas Exchange

3.2.8Lung Disease

3.2.9Lung Disease Data

3.2.10End of Topic Test - Gas Exchange

3.2.11A-A* (AO3/4) - Gas Exchange

3.3Digestion & Absorption

3.3.1Overview of Digestion

3.3.2Digestion in Mammals

3.3.3Absorption

3.3.4End of Topic Test - Substance Exchange & Digestion

3.3.5A-A* (AO3/4) - Substance Ex & Digestion

3.4Mass Transport

3.4.1Haemoglobin

3.4.2Oxygen Transport

3.4.3The Circulatory System

3.4.4The Heart

3.4.5Blood Vessels

3.4.6Cardiovascular Disease

3.4.7Heart Dissection

3.4.8Xylem

3.4.9Phloem

3.4.10Investigating Plant Transport

3.4.11End of Topic Test - Mass Transport

3.4.12A-A* (AO3/4) - Mass Transport

4Genetic Information & Variation

4.1DNA, Genes & Chromosomes

4.1.1DNA

4.1.2Genes

4.1.3A-A* (AO3/4) - DNA

4.2DNA & Protein Synthesis

4.2.1Protein Synthesis

4.2.2Transcription & Translation

4.2.3End of Topic Test - DNA, Genes & Protein Synthesis

4.2.4Exam-Style Question - Protein Synthesis

4.2.5A-A* (AO3/4) - Coronavirus Translation

4.2.6A-A* (AO3/4) - Transcription

4.2.7A-A* (AO3/4) - Translation

4.3Mutations & Meiosis

4.3.1Mutations

4.3.2Meiosis

4.3.3A-A* (AO3/4) - Meiosis

4.3.4Meiosis vs Mitosis

4.3.5End of Topic Test - Mutations, Meiosis

4.3.6A-A* (AO3/4) - DNA,Genes, CellDiv & ProtSynth

4.4Genetic Diversity & Adaptation

4.4.1Genetic Diversity

4.4.2Natural Selection

4.4.3A-A* (AO3/4) - Natural Selection

4.4.4Adaptations

4.4.5Investigating Natural Selection

4.4.6End of Topic Test - Genetic Diversity & Adaptation

4.4.7A-A* (AO3/4) - Genetic Diversity & Adaptation

4.5Species & Taxonomy

4.5.1Classification

4.5.2DNA Technology

4.5.3A-A* (AO3/4) - Species & Taxonomy

4.6Biodiversity Within a Community

4.6.1Biodiversity

4.6.2Agriculture

4.6.3End of Topic Test - Species,Taxonomy& Biodiversity

4.6.4A-A* (AO3/4) - Species,Taxon&Biodiversity

4.7Investigating Diversity

4.7.1Genetic Diversity

4.7.2Quantitative Investigation

5Energy Transfers (A2 only)

6Responding to Change (A2 only)

6.1Nervous Communication

6.1.1Survival

6.1.2Plant Responses

6.1.3Animal Responses

6.1.4Reflexes

6.1.5End of Topic Test - Reflexes, Responses & Survival

6.1.6Receptors

6.1.7The Human Retina

6.1.8Control of Heart Rate

6.1.9End of Topic Test - Receptors, Retina & Heart Rate

6.2Nervous Coordination

6.2.1Neurones

6.2.2Action Potentials

6.2.3Speed of Transmission

6.2.4End of Topic Test - Neurones & Action Potentials

6.2.5Synapses

6.2.6Types of Synapse

6.2.7Medical Application

6.2.8End of Topic Test - Synapses

6.2.9A-A* (AO3/4) - Nervous Comm&Coord

6.3Muscle Contraction

6.3.1Skeletal Muscle

6.3.2Sliding Filament Theory

6.3.3Contraction

6.3.4Slow & Fast Twitch Fibres

6.3.5End of Topic Test - Muscles

6.3.6A-A* (AO3/4) - Muscle Contraction

6.4Homeostasis

6.4.1Overview of Homeostasis

6.4.2Blood Glucose Concentration

6.4.3Controlling Blood Glucose Concentration

6.4.4End of Topic Test - Blood Glucose

6.4.5Primary & Secondary Messengers

6.4.6Diabetes Mellitus

6.4.7Measuring Glucose Concentration

6.4.8Osmoregulation

6.4.9Controlling Blood Water Potential

6.4.10ADH

6.4.11End of Topic Test - Diabetes & Osmoregulation

6.4.12A-A* (AO3/4) - Homeostasis

7Genetics & Ecosystems (A2 only)

7.1Genetics

7.1.1Key Terms in Genetics

7.1.2Inheritance

7.1.3Linkage

7.1.4Multiple Alleles & Epistasis

7.1.5Chi-Squared Test

7.1.6End of Topic Test - Genetics

7.1.7A-A* (AO3/4) - Genetics

7.2Populations

7.2.1Populations

7.2.2Hardy-Weinberg Principle

7.3Evolution

7.3.1Variation

7.3.2Natural Selection & Evolution

7.3.3End of Topic Test - Populations & Evolution

7.3.4Types of Selection

7.3.5Types of Selection Summary

7.3.6Overview of Speciation

7.3.7Causes of Speciation

7.3.8Diversity

7.3.9End of Topic Test - Selection & Speciation

7.3.10A-A* (AO3/4) - Populations & Evolution

7.4Populations in Ecosystems

7.4.1Overview of Ecosystems

7.4.2Niche

7.4.3Population Size

7.4.4Investigating Population Size

7.4.5End of Topic Test - Ecosystems & Population Size

7.4.6Succession

7.4.7Conservation

7.4.8End of Topic Test - Succession & Conservation

7.4.9A-A* (AO3/4) - Ecosystems

8The Control of Gene Expression (A2 only)

8.1Mutation

8.1.1Mutations

8.1.2Effects of Mutations

8.1.3Causes of Mutations

8.2Gene Expression

8.2.1Stem Cells

8.2.2Stem Cells in Disease

8.2.3End of Topic Test - Mutation & Gene Epression

8.2.4A-A* (AO3/4) - Mutation & Stem Cells

8.2.5Regulating Transcription

8.2.6Epigenetics

8.2.7Epigenetics & Disease

8.2.8Regulating Translation

8.2.9Experimental Data

8.2.10End of Topic Test - Transcription & Translation

8.2.11Tumours

8.2.12Correlations & Causes

8.2.13Prevention & Treatment

8.2.14End of Topic Test - Cancer

8.2.15A-A* (AO3/4) - Gene Expression & Cancer

8.3Genome Projects

8.3.1Using Genome Projects

8.4Gene Technology

8.4.1Recombinant DNA

8.4.2Producing Fragments

8.4.3Amplification

8.4.4End of Topic Test - Genome Project & Amplification

8.4.5Using Recombinant DNA

8.4.6Medical Diagnosis

8.4.7Genetic Fingerprinting

8.4.8End of Topic Test - Gene Technologies

8.4.9A-A* (AO3/4) - Gene Technology

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