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.

1Unity & Diversity - Molecules

1.1Water

1.1.1Water & Hydrogen Bonding

1.1.2IB Multiple Choice - Water & Hydrogen Bonding

1.1.3Extended Response - Water

1.2DNA Structure & Replication

1.2.1DNA

1.2.2RNA

1.2.3DNA Replication

1.2.4DNA Replication - Summary

1.2.5IB Multiple Choice - DNA Replication

1.3Transcription & Gene Expression

1.3.1Introduction to Transcription & Protein Synthesis

1.3.2Types of RNA

1.3.3Transcription

1.3.4RNA Processing in Eukaryotes

1.3.5IB Multiple Choice - Transcription

1.4Translation

1.4.1Translation

1.4.2Retroviruses

1.4.3Scientific Practices: Coronavirus

1.4.4IB Multiple Choice - Translation

2Unity & Diversity - Cells

2.1The Origin of Cells

2.1.1Origins - Endosymbiosis

2.2Introduction to Cells

2.2.1Introduction to Cells

2.2.2Cell Size & Surface Area

2.2.3Surface Area to Volume Ratio

2.2.4Eukaryotic vs Prokaryotic Cells

2.2.5Stem Cells

2.2.6Stem Cells in Disease

2.3Ultrastructure of Cells

2.3.1Golgi, Lysosomes & Ribosomes

2.3.2Cell Walls & Vacuole

2.3.3Prokaryotes

2.3.4Binary Fission

2.4Cell Division

2.4.1The Cell Cycle

2.4.2Mitosis

2.4.3Regulation of the Cell Cycle

2.4.4End of Topic Quiz - The Cell Cycle

2.4.5IB Multiple Choice - The Cell Cycle

2.4.6Extended Response - Mitosis

2.5Structure of DNA & RNA

2.5.1DNA & RNA

2.6DNA Replication, Transcription & Translation

2.6.1Protein Synthesis

2.6.2Transcription & Translation

2.6.3End of Topic Test - DNA, Genes & Protein Synthesis

2.6.4Exam-Style Question - Protein Synthesis

2.6.5End of Topic Test - Coronavirus Translation

2.6.6IB Multiple Choice - Transcription

2.6.7IB Multiple Choice - Translation

2.7Cell Respiration

2.7.1Overview of Respiration

2.7.2Anaerobic Respiration

2.7.3End of Topic Test - Anaerobic Respiration

2.7.4Respiration Experiments

2.7.5End of Topic Test - Respiration

2.7.6IB Multiple Choice - Respiration

2.8Photosynthesis

2.8.1Overview of Photosynthesis

2.9Viruses

2.9.1Viruses

3Unity & Diversity - Organisms

3.1Diversity of Organisms

3.1.1Chromosomal Theory of Inheritance

3.1.2Chromosomal Basis of Inherited Disorders

3.1.3IB Multiple Choice - Chromosomal Inheritance

3.2Evidence for Evolution

3.2.1Evidence for Evolution - Fossils & DNA

3.2.2Evidence for Evolution - Anatomy & Geography

3.2.3IB Multiple Choice - Evidence for Evolution

3.2.4Extended Response - DNA & Evolution

3.3Natural Selection

3.3.1Introduction

3.3.2Evolutionary Fitness

3.3.3Natural Selection & Variation

3.3.4End of Topic Quiz - Natural Selection

3.3.5IB Multiple Choice - Natural Selection

3.3.6Speciation

4Unity & Diversity - Ecosystems

4.1Classification

4.1.1Introduction to Biodiversity

4.1.2Keystone Species

4.1.3Courtship & Ancestry

4.1.4Classification

4.2Cladistics

4.2.1Evolutionary Relationships

4.2.2IB Multiple Choice - Species & Taxonomy

4.3Evolution & Speciation

4.3.1Evidence for Evolution - Fossils & DNA

4.3.2Evidence for Evolution - Anatomy & Geography

4.3.3IB Multiple Choice - Evidence for Evolution

4.3.4Extended Response - DNA & Evolution

4.3.5Populations

4.3.6Mutations, Genetic Drift, & Gene Flow

4.3.7Speciation

4.3.8Rate of Speciation

4.3.9Allopatric & Sympatric Speciation

4.4Conservation of Biodiversity

4.4.1Coming Soon!

5Form & Function - Molecules

5.1Carbohydrates & Lipids

5.1.1Monosaccharides

5.1.2Disaccharides & Polysaccharides

5.1.3Triglycerides & Phospholipids

5.1.4Cellulose

5.1.5Starch & Glycogen

5.2Proteins

5.2.1Structure & Functions of Proteins

6Form & Function - Cells

6.1Membranes & Membrane Transport

6.1.1Passive Movement Across Membranes

6.1.2Active Movement Across Membranes

6.1.3Endocytosis & Exocystosis

6.1.4Cell Membrane Structure & Permeability

6.2Organelles & Compartmentalization

6.2.1Eukaryotic Cells & Organelles

6.2.2Subcellular Components & Organelles

6.3Cell Specialization

6.3.1Cell Specialization

7Form & Function - Organisms

7.1Gas Exchange

7.1.1Lung Structure & Alveoli

7.1.2Ventilation

7.1.3Lung Disease

7.1.4Lung Disease Data

7.2Transport

7.2.1The Circulatory System

7.2.2The Heart

7.2.3Blood Vessels

7.2.4Cardiovascular Disease

7.2.5Xylem Structure

7.2.6Cohesion-Tension Theory

7.2.7Investigating Transpiration Rate

7.2.8Phloem Structure

7.2.9Translocation

7.3Muscle & Motility

7.3.1Skeletal Muscle

7.3.2Sliding Filament Theory

7.3.3Contraction

8Form & Function - Ecosystems

8.1Species, Communities & Ecosytems

8.1.1Population Dynamics

8.1.2Communities & Measuring Diversity

8.1.3Relationships Between Populations

8.1.4Overview of Ecosystems

8.1.5Nutrient Cycles

8.1.6Fertilisers

8.1.7Eutrophication

8.1.8Chi-Squared Test

8.2Energy Flow

8.2.1Energy

8.2.2Energy Flow

8.2.3Biomass & Food Chains

8.2.4Heterotrophs & Autotrophs

8.3Carbon Cycle

8.3.1Carbon Cycle

8.4Climate Change

8.4.1Carbon Dioxide

8.4.2The Greenhouse Effect

8.4.3Global Warming

8.4.4Maintaining Biodiversity

9Interaction & Interdependence - Molecules

9.1Enzymes

9.1.1Enzymes

9.1.2Factors Affecting Enzyme Activity

9.1.3Enzyme-Controlled Reactions

9.1.4End of Topic Test - Enzymes

9.2Metabolism

9.2.1Molecules to Metabolism

9.3Cell Respiration

9.3.1Glycolysis

9.3.2Aerobic Respiration

9.4Photosynthesis

9.4.1Light-Dependent Reaction

9.4.2Light-Independent Reaction

9.4.3End of Topic Test - Photosynthesis Reactions

9.4.4Limiting Factors

9.4.5Photosynthesis Experiments

9.4.6End of Topic Test - Photosynthesis

9.4.7End of Topic Test - Photosynthesis

10Interaction & Interdependence - Cells

10.1Chemical Signalling

10.1.1Coming Soon!

10.2Neural Signalling

10.2.1Medical Application

10.2.2Speed of Transmission

10.2.3Synapses

10.2.4End of Topic Test - Nervous Comm&Coord

10.2.5Neuron Structure

10.2.6Types of Synapse

10.2.7End of Topic Test - Neurones & Action Potentials

10.2.8Transmission of an Impulse

10.2.9End of Topic Test - Synapses

10.3Adaptation to Environment

10.3.1Coming Soon!

10.4Ecological Niches

10.4.1Coming Soon!

11Interaction & Interdependence - Organisms

11.1Integration of Body Systems

11.1.1Plant Tropisms

11.2Defence Against Disease

11.2.1Immune System

11.2.2The Immune Response

11.2.3Antibodies

11.2.4Primary & Secondary Response

11.2.5Vaccines

11.2.6HIV

11.2.7End of Topic Test - Immune System

12Interaction & Interdependence - Ecosystems

12.1Populations & Communities

12.1.1Coming Soon!

12.2Transfers of Energy & Matter

12.2.1Coming Soon!

13Continuity & Change - Molecules

13.1DNA Replication

13.1.1DNA Replication

13.2Protein Synthesis

13.2.1Protein Synthesis

13.2.2Transcription & Translation

13.2.3End of Topic Test - DNA, Genes & Protein Synthesis

13.3Mutation & Gene Editing

13.3.1Genes

13.3.2Mutations

13.3.3DNA Technology

13.3.4PCR & Gel Electrophoresis

13.3.5Genetic Modification

13.3.6Cloning

14Continuity & Change - Cells

14.1Gene Expression

14.1.1Introduction to Transcription & Protein Synthesis

14.1.2Types of RNA

14.1.3Transcription

14.1.4RNA Processing in Eukaryotes

14.1.5IB Multiple Choice - Transcription

14.2Water Potential

14.2.1Controlling Blood Water Potential

15Continuity & Change - Organisms

15.1Inheritance

15.1.1Non-Nuclear Inheritance

15.1.2Linked Genes

15.1.3IB Multiple Choice - Non-Mendelian Genetics

15.1.4Extended Response - Inheritance

15.1.5Introduction to Non-Mendelian Inheritance

15.1.6Chi-Squared Test

15.1.7End of Topic Quiz - Inheritance

15.1.8Sex-Linked Genes

15.1.9Grade 4-5 (Scientific Practices) - Inheritance

15.2Homeostasis

15.2.1Overview of Homeostasis

15.2.2Blood Glucose Concentration

15.2.3Controlling Blood Glucose Concentration

15.2.4End of Topic Test - Blood Glucose

15.2.5Type I & Type II Diabetes

15.2.6Human Reproductive Systems

15.2.7Hormonal Control of the Menstrual Cycle

15.2.8Kidneys & Osmoregulation

16Continuity & Change - Ecosystems

16.1Natural Selection

16.1.1Natural Selection

16.2Stability & Change

16.2.1Coming Soon!

16.3Climate Change

16.3.1Carbon Dioxide

16.3.2The Greenhouse Effect

16.3.3Global Warming

16.3.4Maintaining Biodiversity

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