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Questions and Answers
Place the beaker set-ups in a water bath and let boil until the pigment is extracted (when leaves are completely devoid of ______)
Place the beaker set-ups in a water bath and let boil until the pigment is extracted (when leaves are completely devoid of ______)
color
Do not boil under direct ______
Do not boil under direct ______
flame
Always boil in a water ______
Always boil in a water ______
bath
Remove the leaves from the alcohol and spread them out in a ______ dish
Remove the leaves from the alcohol and spread them out in a ______ dish
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Add just enough iodine solution to cover the ______
Add just enough iodine solution to cover the ______
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The presence of starch is indicated by a bluish black ______
The presence of starch is indicated by a bluish black ______
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Photosynthesis is the process whereby green plants convert the radiant energy of light into chemical energy that is then stored in ______ molecules.
Photosynthesis is the process whereby green plants convert the radiant energy of light into chemical energy that is then stored in ______ molecules.
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For photosynthesis to occur, light, a green pigment and ______ dioxide must be present.
For photosynthesis to occur, light, a green pigment and ______ dioxide must be present.
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All living things are dependent on the products of photosynthesis for their energy, for the raw materials necessary for growth and repair and for the ______ necessary for life.
All living things are dependent on the products of photosynthesis for their energy, for the raw materials necessary for growth and repair and for the ______ necessary for life.
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The process of photosynthesis takes place in ______.
The process of photosynthesis takes place in ______.
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Glucose, the main product of photosynthesis, can be converted into starch, a storage molecule that can be broken down to glucose again when the plant requires it, during ______ respiration.
Glucose, the main product of photosynthesis, can be converted into starch, a storage molecule that can be broken down to glucose again when the plant requires it, during ______ respiration.
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At the end of the exercise, the student is expected to be able to measure the rate of photosynthesis as affected by some factors and detect the presence or absence of photosynthetic products in leaves, and extract and identify the pigments involved in ______.
At the end of the exercise, the student is expected to be able to measure the rate of photosynthesis as affected by some factors and detect the presence or absence of photosynthetic products in leaves, and extract and identify the pigments involved in ______.
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Set the three test tubes in bright ______.
Set the three test tubes in bright ______.
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Chromatography is a method of separating closely related compounds by allowing the mixture of substances to diffuse through an absorbent called ______.
Chromatography is a method of separating closely related compounds by allowing the mixture of substances to diffuse through an absorbent called ______.
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The different chloroplast pigments can be separated and observed as different color bands on the ______.
The different chloroplast pigments can be separated and observed as different color bands on the ______.
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Chlorophyll pigments can be identified not just by their band color but also by the distance they travel through the ______.
Chlorophyll pigments can be identified not just by their band color but also by the distance they travel through the ______.
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The Rf (retention factor) value is a ratio that can be calculated and compared to known Rf standards to identify unknown ______.
The Rf (retention factor) value is a ratio that can be calculated and compared to known Rf standards to identify unknown ______.
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Grind the leaves using a pestle to release the ______.
Grind the leaves using a pestle to release the ______.
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Add another 3 ml of ______ and continue crushing until the acetone becomes a very dark, highly concentrated extract.
Add another 3 ml of ______ and continue crushing until the acetone becomes a very dark, highly concentrated extract.
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Collect the extract by pouring the solution into a beaker covered with ______.
Collect the extract by pouring the solution into a beaker covered with ______.
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Fold the cheesecloth and squeeze the extract to release all the liquid into the beaker. Cover the beaker when ______.
Fold the cheesecloth and squeeze the extract to release all the liquid into the beaker. Cover the beaker when ______.
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Cut a strip of filter paper about 1 inch wide and long enough to reach the solvent at the bottom of the beaker that you will use as your chromatography “tank”. Cut the filter paper to the shape shown in Figure 1. This is referred to as a ______.
Cut a strip of filter paper about 1 inch wide and long enough to reach the solvent at the bottom of the beaker that you will use as your chromatography “tank”. Cut the filter paper to the shape shown in Figure 1. This is referred to as a ______.
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Using a pencil, draw a faint line 1 cm from the pointed tip. This marks your solvent’s origin. With the capillary tube/pipette, apply three to five drops of the pigment extract just above the ______ line.
Using a pencil, draw a faint line 1 cm from the pointed tip. This marks your solvent’s origin. With the capillary tube/pipette, apply three to five drops of the pigment extract just above the ______ line.
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Pour the prepared separating fluid consisting of a 1:9 mixture of acetone and petroleum ether into the chromatography tank (tall beaker) to a depth of a little more than 0.5 cm. ______
Pour the prepared separating fluid consisting of a 1:9 mixture of acetone and petroleum ether into the chromatography tank (tall beaker) to a depth of a little more than 0.5 cm. ______
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Carefully lower the extract-treated chromatogram into the tank, with the end immersed in the separating fluid but not reaching the level of the pigment spot. Secure the chromatogram with a ______.
Carefully lower the extract-treated chromatogram into the tank, with the end immersed in the separating fluid but not reaching the level of the pigment spot. Secure the chromatogram with a ______.
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Make sure that the chromatogram is suspended freely, with no sides touching the tank or other chromatograms. The solvent will move up the strip and carry the pigments with it. Observe the set-up around a 15-to-30-minute period as the separating fluid diffuses upward along the length of the chromatogram, passing through the spot and gradually separating the chloroplast pigments. Draw the separated pigments on your worksheet and label each color band by its corresponding ______.
Make sure that the chromatogram is suspended freely, with no sides touching the tank or other chromatograms. The solvent will move up the strip and carry the pigments with it. Observe the set-up around a 15-to-30-minute period as the separating fluid diffuses upward along the length of the chromatogram, passing through the spot and gradually separating the chloroplast pigments. Draw the separated pigments on your worksheet and label each color band by its corresponding ______.
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Using a ruler, measure the distance from the origin line to the solvent front (visible as a thin, transparent line at the top of the chromatogram) and to the middle section of each color band (at its widest section). Calculate the ______ values for each pigment.
Using a ruler, measure the distance from the origin line to the solvent front (visible as a thin, transparent line at the top of the chromatogram) and to the middle section of each color band (at its widest section). Calculate the ______ values for each pigment.
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Fill in the table in your worksheet. Compare your ______ values with the known standards of each pigment for the specific solvent as follows: a.Carotene – 0.95 b.Phaeophytin – 0.83 c.Xanthophyll – 0.71 d.Chlorophyll a – 0.65 e.Chlorophyll b – 0.45 FORMATIVE LEARNING ACTIVITY: Labster: Refer to the Labster link in your canvas for the online simulations Simulation: Pigment Extraction: Use Photosynthesis to Produce Biofuel Simulation: Algae Pigment Analysis Adapted from: De Jesus J, Ragragio E. 2000.Laboratory Manual in General Botany.
Fill in the table in your worksheet. Compare your ______ values with the known standards of each pigment for the specific solvent as follows: a.Carotene – 0.95 b.Phaeophytin – 0.83 c.Xanthophyll – 0.71 d.Chlorophyll a – 0.65 e.Chlorophyll b – 0.45 FORMATIVE LEARNING ACTIVITY: Labster: Refer to the Labster link in your canvas for the online simulations Simulation: Pigment Extraction: Use Photosynthesis to Produce Biofuel Simulation: Algae Pigment Analysis Adapted from: De Jesus J, Ragragio E. 2000.Laboratory Manual in General Botany.
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Carefully lower the extract-treated chromatogram into the tank, with the end immersed in the separating fluid but not reaching the level of the pigment spot. Secure the chromatogram with a paper clip. Make sure that the chromatogram is suspended freely, with no sides touching the tank or other chromatograms. The solvent will move up the strip and carry the pigments with it. Observe the set-up around a 15-to-30-minute period as the separating fluid diffuses upward along the length of the chromatogram, passing through the spot and gradually separating the chloroplast pigments. Draw the separated pigments on your worksheet and label each color band by its corresponding pigment. Using a ruler, measure the distance from the origin line to the solvent front (visible as a thin, transparent line at the top of the chromatogram) and to the middle section of each color band (at its widest section). Calculate the Rf values for each pigment. Fill in the table in your worksheet. Compare your Rf values with the known standards of each pigment for the specific solvent as follows: a.Carotene – 0.95 b.Phaeophytin – 0.83 c.Xanthophyll – 0.71 d.Chlorophyll a – 0.65 e.Chlorophyll b – 0.45 FORMATIVE LEARNING ACTIVITY: Labster: Refer to the Labster link in your canvas for the online simulations Simulation: Pigment Extraction: Use Photosynthesis to Produce Biofuel Simulation: Algae Pigment Analysis Adapted from: De Jesus J, Ragragio E. 2000.Laboratory Manual in General Botany. This process is typically used in ______ analysis.
Carefully lower the extract-treated chromatogram into the tank, with the end immersed in the separating fluid but not reaching the level of the pigment spot. Secure the chromatogram with a paper clip. Make sure that the chromatogram is suspended freely, with no sides touching the tank or other chromatograms. The solvent will move up the strip and carry the pigments with it. Observe the set-up around a 15-to-30-minute period as the separating fluid diffuses upward along the length of the chromatogram, passing through the spot and gradually separating the chloroplast pigments. Draw the separated pigments on your worksheet and label each color band by its corresponding pigment. Using a ruler, measure the distance from the origin line to the solvent front (visible as a thin, transparent line at the top of the chromatogram) and to the middle section of each color band (at its widest section). Calculate the Rf values for each pigment. Fill in the table in your worksheet. Compare your Rf values with the known standards of each pigment for the specific solvent as follows: a.Carotene – 0.95 b.Phaeophytin – 0.83 c.Xanthophyll – 0.71 d.Chlorophyll a – 0.65 e.Chlorophyll b – 0.45 FORMATIVE LEARNING ACTIVITY: Labster: Refer to the Labster link in your canvas for the online simulations Simulation: Pigment Extraction: Use Photosynthesis to Produce Biofuel Simulation: Algae Pigment Analysis Adapted from: De Jesus J, Ragragio E. 2000.Laboratory Manual in General Botany. This process is typically used in ______ analysis.
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Carefully lower the extract-treated chromatogram into the tank, with the end immersed in the separating fluid but not reaching the level of the pigment spot. Secure the chromatogram with a paper clip. Make sure that the chromatogram is suspended freely, with no sides touching the tank or other chromatograms. The solvent will move up the strip and carry the pigments with it. Observe the set-up around a 15-to-30-minute period as the separating fluid diffuses upward along the length of the chromatogram, passing through the spot and gradually separating the chloroplast pigments. Draw the separated pigments on your worksheet and label each color band by its corresponding pigment. Using a ruler, measure the distance from the origin line to the solvent front (visible as a thin, transparent line at the top of the chromatogram) and to the middle section of each color band (at its widest section). Calculate the Rf values for each pigment. Fill in the table in your worksheet. Compare your Rf values with the known standards of each pigment for the specific solvent as follows: a.Carotene – 0.95 b.Phaeophytin – 0.83 c.Xanthophyll – 0.71 d.Chlorophyll a – 0.65 e.Chlorophyll b – 0.45 FORMATIVE LEARNING ACTIVITY: Labster: Refer to the Labster link in your canvas for the online simulations Simulation: Pigment Extraction: Use Photosynthesis to Produce Biofuel Simulation: Algae Pigment Analysis Adapted from: De Jesus J, Ragragio E. 2000.Laboratory Manual in General Botany. The Rf value represents the ratio of the distance traveled by the ______ to the distance traveled by the solvent front.
Carefully lower the extract-treated chromatogram into the tank, with the end immersed in the separating fluid but not reaching the level of the pigment spot. Secure the chromatogram with a paper clip. Make sure that the chromatogram is suspended freely, with no sides touching the tank or other chromatograms. The solvent will move up the strip and carry the pigments with it. Observe the set-up around a 15-to-30-minute period as the separating fluid diffuses upward along the length of the chromatogram, passing through the spot and gradually separating the chloroplast pigments. Draw the separated pigments on your worksheet and label each color band by its corresponding pigment. Using a ruler, measure the distance from the origin line to the solvent front (visible as a thin, transparent line at the top of the chromatogram) and to the middle section of each color band (at its widest section). Calculate the Rf values for each pigment. Fill in the table in your worksheet. Compare your Rf values with the known standards of each pigment for the specific solvent as follows: a.Carotene – 0.95 b.Phaeophytin – 0.83 c.Xanthophyll – 0.71 d.Chlorophyll a – 0.65 e.Chlorophyll b – 0.45 FORMATIVE LEARNING ACTIVITY: Labster: Refer to the Labster link in your canvas for the online simulations Simulation: Pigment Extraction: Use Photosynthesis to Produce Biofuel Simulation: Algae Pigment Analysis Adapted from: De Jesus J, Ragragio E. 2000.Laboratory Manual in General Botany. The Rf value represents the ratio of the distance traveled by the ______ to the distance traveled by the solvent front.
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Study Notes
Photosynthesis
- Photosynthesis is the process of converting light energy into chemical energy, which is stored in food molecules.
- It requires light, a green pigment, and carbon dioxide to occur.
- All living things, except some microorganisms, rely on photosynthesis for energy, growth, and repair.
Photosynthesis Equation
- The basic equation for photosynthesis is: 6CO2 + 12H2O → C6H12O6 + 6H2O + 6O2↑
- Glucose, the main product of photosynthesis, has three different pathways:
- Conversion into other chemical substances required by plant cells
- Conversion into starch for storage
- Breakdown during cellular respiration to release energy
Objectives
- Measure the rate of photosynthesis affected by various factors
- Detect the presence or absence of photosynthetic products in leaves
- Extract and identify the pigments involved in photosynthesis
Experiment: Detecting Starch
- Boil leaves in a water bath with ethyl alcohol to extract pigments
- Add iodine solution to the leaves to detect starch presence
- The presence of starch is indicated by a bluish-black color
Experiment: Intensity of Light
- Observe the effect of light intensity on photosynthesis
- Results show that increasing light intensity increases photosynthesis rate
Experiment: Wavelength of Light
- Study the effect of different wavelengths of light on photosynthesis
- Data shows that photosynthesis occurs at different rates depending on the wavelength of light
Experiment: Carbon Dioxide Requirement
- Investigate the role of carbon dioxide in photosynthesis
- Results show that carbon dioxide is essential for photosynthesis
Chromatography
- Chromatography is a method for separating and identifying closely related compounds, such as pigments
- The distance a pigment travels through the chromatogram, known as the Rf value, is characteristic of the compound and solvent used
Pigment Extraction and Chromatography
- Extract pigments from green and variegated leaves using acetone
- Separate the pigments using paper chromatography
- Identify the pigments based on their color bands and Rf values
- Calculate the Rf values and compare them to known standards for each pigment
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Description
Learn about the process of separating chloroplast pigments using paper chromatography. Observe the color changes in test tubes and document the observations in a worksheet. Understand how chromatography is used to differentiate closely related compounds like pigments.
