Chromatography of Plant Pigments 3

 

 

Chromatography of Plant Pigments

 

 

Introduction:

 

Can chromatography be used to separate mixtures of chemical substances? The purpose of this experiment is to answer this question. In paper chromatography, a liquid sample flows down a vertical strip of absorbent paper, on which the components of a mixture are deposited in specific directions and locations. Chromatography is a tool used to examine and separate mixtures of chemical substances. Chromatography is essential to the separation of pure substances from complex mixtures. Separation results in a chromatographically pure substance. Chromatography allows you to determine the properties of chemical substances.

The relationship between the chromatography paper, mixture, and the solvent is very important in all chromatographic separations. The solvent has to dissolve the mixture that should be separated. The paper must also absorb the components of the mixtures selectively and reversibly. The substances making up the mixture must be evenly dispersed in the water. Chromatography is a simple and inexpensive tool for separating and identifying chemical mixtures if all these things are done.

 

Hypothesis:

 

Paper can be used to separate mixed chemicals.

 

Materials:

 

The materials used in this lab are filter paper, test tube, rubber stopper, paper clip, metric ruler, black felt-tip pen, pencil, calculator, and water.

 

Methods:

 

First, bend a paper clip so that it’s straight with a hook at one end. Push the straight end of the paper clip into the bottom of a cork stopper. Then, hang a thin strip of filter paper on the hooked end of the paper clip and insert the paper strip into the test tube. The paper should not touch the sides and should almost touch the bottom of the test tube. Next, remove the paper strip from the test tube. Now draw a solid 5-mm-wide band about 25 mm from the bottom of the paper, using a black felt tip pen. After this, use a pencil to draw a line across the paper strip 10 cm above the black band. Then, put the filter paper back into the test tube with the bottom of the paper in the water and the black band above the water. Observe what happens as the liquid travels up the paper and record the changes you see. When the solvent has reached the pencil line, remove the paper from the test tube. Let the paper dry on the desk. Finally, with a metric ruler, measure the distances from the starting point to the top edge of each color. Record the data in a data table and calculate a ratio for each color by dividing the distance, the color traveled by the distance the solvent traveled.

 

Results:

 

The results of the chromatography experiment are shown in a chart and a graph.

 

Color of ink (list in order) Distance traveled by each color (mm) Distance solvent traveled (mm) Ratio traveled = distance color moved divided by distance solvent moved
Yellow 70 108 0.65
Orange  

85

 

108

 

0.79

Pink 95  

108

 

0.88

Violet 102  

108

 

0.94

Blue 108  

108

 

1.00

 

 

 

Questions:

1. How many colors separated from the black ink? Five colors separated from the ink: yellow, orange, pink, violet, and blue.

 

2. What served as the solvent for the ink? Water served as the solvent for the ink.

 

3. As the solvent traveled up the paper, which color of ink appeared first? Dark blue appeared first.

 

4. List the colors in order from top to bottom that separated from the black ink? The colors separated in the order of: blue, violet, pink, orange, and yellow.

 

5. In millimeters, how far did the solvent travel? The solvent traveled 108 mm.

 

6. From your results, what can you conclude is true about black ink? Black ink is a mixture of several different colors.

 

7. Why did the inks separate? The inks separated because black ink is a mixture of different pigments that are soluble in water, have different molecular characteristics, and travel different distances.

 

8. Why did some inks move a greater distance? Some inks move a greater distance because molecules in ink have different characteristics, like how readily they are absorbed by paper. This means that the ink least readily absorbed by paper will travel farthest from the starting mark and the ink most readily absorbed by paper will be the closest to the starting mark. All of the different color inks that were separated were different in how readily they are absorbed by paper.

Error Analysis:

 

There are a few errors that could have changed the results. First, there could be inaccurate measurements of how far every color traveled or how far the water traveled up the filter paper. Another error could occur when calculating the ratio traveled, Rf value. Also, a longer test tube could have been used by different groups which would make the filter strip longer. This means that a group could have detected another color because they had more room on their filter paper. This also could have affected the ratios. Finally, the groups could have put different amounts of black ink on the filter paper.

 

Conclusion:

 

The hypothesis that paper can be used to separate mixed chemicals was correct. The different colored inks mixed together give the black its color. The five colors that separated from the black ink were blue, violet, pink, orange, and yellow. Blue appeared first and then was followed by violet, pink, orange, and yellow. The colors separated the way they did because they have different molecular characteristics, like how readily they were absorbed by the paper and their solubility in water. Blue was most readily absorbed by the paper and soluble by water, while yellow was the least.

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Chromatography of Simulated Plant Pigments

 

Chromatography of Simulated Plant Pigments

 

Introduction
    This experiment is conducted to investigate the components Plant Pigments separating visibly. There are a couple of different types of components in plant pigments, and they became clearly visible during this lab. The most important and abundant chemical pigment found in plants is chlorophyll. This pigment exists in two forms; chlorophyll a and chlorophyll b. Chlorophyll absorbs two main colors from light quite well. These are blue, and red. The chlorophyll reflects green light very well, however, the two different types of chlorophyll have their maximum absorption at different wavelengths of light. Chlorophyll a, being the main photosynthetic pigment, has a primary purpose to convert light energy to chemical energy used by the plant itself. Chlorophyll b absorbs light in a region of the spectrum apart from the dominant chlorophyll, and transfers the energy it produces to chlorophyll a. Along with chlorophyll b in transferring their energy produced to the dominant chlorophyll, two other pigments that are found in plants are carotenes and xanthophylls, which are orange and yellow respectively. Since chlorophyll is such a dominant pigment in green plants, this domination hides the color of the carotenes and xanthophylls in the leaves. This causes most plant leaves to appear green most of the time. During the autumn, however, the chlorophyll starts to break down, causing the carotenes and xanthophylls to show their bright red, orange and yellow colors.
These brilliant colors can be separated another way. This different technique, known as paper chromatography, separates mixtures in a liquid into individual components. The technique is based on the fact that each substance in a mixture has a specific affinity for a solid surface and a specific solubility in different solvents. By this method, the solid surface is the cellulose fibers in the chromatography paper, and the solvent is the solution that was placed in the bottom of the developing chamber.
This separation takes place through a process of absorption and capillary action. Just a small drop of the mixture, in this case plant pigment to be separated, is placed at the bottom of the strip of chromatography paper. The chromatography paper is then placed in the developing chamber with a solvent, which wicks up the paper, pulling the solvent up the paper by capillary action, and the mixture of pigments is dissolved as the solvent passes over it. The different components of the mixture move upward at different rates. A compound with greater solubility will travel farther than one with less solubility. The pigments then show up as color streaks on the chromatography paper. These substances have formed a pattern called a chromatogram on the chromatography paper.
The Rf values for each pigment is calculated to establish the relative rate of migration for each pigment. This value represents the ratio of the distance a pigment traveled on the chromatogram relative to the distance the solvent front moved.
Scientists use the Rf value of a sample to identify the molecule. Any molecule in a given solvent matrix system has a uniquely consistent Rf value. The formula for this value is as follows:

Rf = Distance each pigment traveled ¸ Distance solvent front traveled

 

Hypothesis
    Using paper chromatography, the pigments that give a leaf its color can be separated and observed to determine the Rf value of each pigment and their function during photosynthesis.

 

Materials
For this experiment the following items are used — one chromatography reaction chamber, one paper chromatography strip, one capillary pipette, a pencil and paper, calculator, ruler, 50 ml beaker, colored pencils, approximately 10 ml of solvent depending on the size of the reaction chamber, scissors, and simulated plant pigment.

 

Procedure
Use scissors to cut the bottom of the chromatography paper to a tapered end. Measure the strip and cut the length to equal slightly longer than the reaction chamber. Draw a faint pencil line at the bottom of the tapered end and use a capillary pipette to add some simulated plant pigment to this line. Add 5-10 ml of solvent to the reaction chamber. Extend the chromatography strip through the slit in the lids of the reaction chamber and carefully lower the strip into the chamber so the tapered end is in the solvent and the pencil line is above the solvent level. Make sure the strip does not touch the walls of the chamber and do not bump the chamber as the pigments begin to separate. After the pigments have completely separated and the solvent front has reached the top of the chamber, remove the strip and mark the solvent front with a pencil line before it evaporates. Measure and record the distance the solvent and each pigment traveled. Use a calculator to determine the Rf values for each pigment.

 

Data

 

Table 1

Band # Pigment Color Migration distance (mm) Rf value
1 Carotene Orange 59mm .94
2 Xanthophyll Yellow 56mm .89
3 Chlorophyll a Light green 29mm .46
4 Chlorophyll b Dark green 14mm .22
Solvent 63mm

Questions
1. Describe what happened to the original spot of simulated plant pigments?
  The solvent separated  the original spot by wicking up the solvent while dissolving the various pigments in the spot.
2. List some other uses of chromatography?  Chromatography can be used to separate various mixtures of subtances, liquids and gases.
3. Which of the 4 pigments migrated the furthest and why?  carotene ( orange) because it was the most soluble in the solvent
4. Which type of chlorophyll was the most soluble?  chlorophyll a
5. Explain why leaves change color in the fall?  In Autumn, chlorophyll starts to break down which allows the other brilliant plant pigment colors to show. These pigments include the red, orange, and yellow colors.
6. What is the function of plant pigments in photosynthesis?  Plant pigments trap light energy and convert it into chemical energy that can be used by the plant to make glucose or sugar.

Error Analysis
The chromatography paper touched the sides of the chamber during the waiting time which caused the migration to go slightly to the side instead of straight to the top. Also the strip was bent at the top so there could have been a slight error in measuring the migration of the solvent  front.

Conclusion
Paper chromatography proved to be an accurate method of separating and observing the various colors of plant pigments. The pigments dissolved in the solvent and migrated upward. The colors were observed and their migration distances measured & recorded. The
Rf value of each pigment was determined by dividing its migration by the migration of the solvent.  It was determined that 4 pigments were present in the original spot — carotene, xanthophyll, chlorophyll a, and chlorophyll b. Carotene was the most soluble, while chlorophyll b was the least soluble.

Chapter 37 AP Objectives

 

Chapter 37     Nutrition in Plants
Objectives
Nutritional Requirements of Plants
1. Describe the ecological role of plants in transforming inorganic molecules into organic compounds.
2. Define the term essential nutrient.
3. Explain how hydroponic culture is used to determine which minerals are essential nutrients.
4. Distinguish between macronutrient and micronutrient.
5. Name the nine macronutrients required by plants.
6. List the eight micronutrients required by plants and explain why plants need only minute quantities of these elements.
7. Explain how a nutrient’s role and mobility determine the symptoms of a mineral deficiency.
The Role of Soil in Plant Nutrition
8. Define soil texture and soil composition.
9. Explain how soil is formed.
10. Name the components of topsoil.
11. Describe the composition of loams and explain why they are the most fertile soils.
12. Explain how humus contributes to the texture and composition of soils.
13. Explain why plants cannot extract all of the water in soil.
14. Explain how the presence of clay in soil helps prevent the leaching of mineral cations.
15. Define cation exchange, explain why it is necessary for plant nutrition, and describe how plants can stimulate the process.
16. Explain why soil management is necessary in agricultural systems but not in natural ecosystems such as forests and grasslands. Describe an example of human mismanagement of soil.
17. List the three mineral elements that are most commonly deficient in agricultural soils.
18. Explain how soil pH determines the effectiveness of fertilizers and a plant’s ability to absorb specific mineral nutrients.
19. Describe problems resulting from farm irrigation in arid regions.
20. Describe actions that can reduce loss of topsoil due to erosion.
21. Explain how phytoremediation can help detoxify polluted soil.
The Special Case of Nitrogen as a Plant Nutrient
22. Define nitrogen fixation and write an overall equation representing the conversion of gaseous nitrogen to ammonia.
23. Explain the importance of nitrogen-fixing bacteria to life on Earth.
24. Summarize the ecological role of each of the following groups of bacteria.
a. ammonifying bacteria
b. denitrifying bacteria
c. nitrogen-fixing bacteria
d. nitrifying bacteria
25. Explain why improving the protein yield of crops is a major goal of agricultural research.
Nutritional Adaptations: Symbiosis of Plants and Soil Microbes
26. Describe the development of a root nodule in a legume.
27. Explain how a legume protects its nitrogen-fixing bacteria from free oxygen, and explain why this protection is necessary.
28. Describe the basis for crop rotation.
29. Explain why a symbiosis between a legume and its nitrogen-fixing bacteria is considered to be mutualistic.
30. Explain why a symbiosis between a plant and a mycorrhizal fungus is considered to be mutualistic.
31. Distinguish between ectomycorrhizae and endomycorrhizae.
Nutritional Adaptations: Parasitism and Predation by Plants
32. Name one modification for nutrition in each of the following groups of plants:
a. epiphytes
b. parasitic plants
c. carnivorous plants
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Chapter 38 AP Objectives

Chapter 38     Plant reproduction and Development
Objectives
Sexual Reproduction
1. In general terms, explain how the basic plant life cycle with alternation of generations is modified in angiosperms.
2. List four floral parts in order from outside to inside a flower.
3. From a diagram of an idealized flower, correctly label the following structures and describe the function of each structure:
a. sepals
b. petals
c. stamen (filament and anther)
d. carpel (style, ovary, ovule, and stigma)
4. Distinguish between:
a. complete and incomplete flowers
b. bisexual and unisexual flowers
c. monoecious and dioecious plant species
5. Explain by which generation, structure, and process spores are produced.
6. Explain by which generation, structure, and process gametes are produced.
7. Name the structures that represent the male and female gametophytes of flowering plants.
8. Describe the development of an embryo sac and explain the fate of each of its cells.
9. Explain how pollen can be transferred between flowers.
10. Distinguish between pollination and fertilization.
11. Describe mechanisms that prevent self-pollination.
12. Outline the process of double fertilization. Explain the adaptive advantage of double fertilization in angiosperms.
13. Explain how fertilization in animals is similar to that in plants.
14. Describe the fate of the ovule and ovary after double fertilization. Note where major nutrients are stored as the embryo develops.
15. Describe the development and function of the endosperm. Distinguish between liquid endosperm and solid endosperm.
16. Describe the development of a plant embryo from the first mitotic division to the embryonic plant with rudimentary organs.
17. From a diagram, identify the following structures of a seed and state a function for each:
a. seed coat
b. proembryo
c. suspensor
d. hypocotyls
e. radicle
f. epicotyl
g. plumule
h. endosperm
i. cotyledons
j. shoot apex
18. Explain how a monocot and dicot seed differ.
19. Explain how fruit forms and ripens.
20. Distinguish among simple, aggregate, and multiple fruit. Give an example of each type of fruit.
21. Explain how selective breeding by humans has changed fruits.
22. Explain how seed dormancy can be advantageous to a plant. Describe some conditions for breaking dormancy.
23. Describe the process of germination in a garden bean.
Asexual Reproduction
24. Describe the natural mechanisms of vegetative reproduction in plants, including fragmentation and apomixis.
25. Explain the advantages and disadvantages of reproducing sexually and asexually.
26. Explain various methods that horticulturalists use to propagate plants from cuttings.
27. Explain how the technique of plant tissue culture can be used to clone and genetically engineer plants.
28. Describe the process of protoplast fusion and its potential agricultural impact.
Plant Biotechnology
29. Compare traditional plant-breeding techniques and genetic engineering, noting similarities and differences.
30. Describe two transgenic crops.
31. Describe some of the biological arguments for and against genetically modified crops.
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Chapter 29 AP Objectives

 

Chapter 29     Plant Diversity I: Colonization of Land
Objectives
An Overview of Land Plant Evolution
1. Describe four shared derived homologies that link charophyceans and land plants.
2. Distinguish among the kingdoms Plantae, Streptophyta, and Viridiplantae. Note which of these is used in the textbook.
3. Describe five characteristics that distinguish land plants from charophycean algae. Explain how these features are adaptive for life on land.
4. Define and distinguish among the stages of the alternation of generations life cycle
5. Describe evidence that suggests that plants arose roughly 475 million years ago.
Bryophytes
6. List and distinguish among the three phyla of bryophytes. Briefly describe the characteristics of each group.
7. Distinguish between the phylum Bryophyta and the bryophytes.
8. Explain why bryophyte rhizoids are not considered roots.
9. Explain why most bryophytes grow close to the ground.
10. Diagram the life cycle of a bryophyte. Label the gametophyte and sporophyte stages and the locations of gamete production, fertilization, and spore production.
11. Describe the ecological and economic significance of bryophytes.
The Origin and Diversity of Vascular Plants
12. Describe the five traits that characterize modern vascular plants. Explain how these characteristics have contributed to their success on land.
13. Distinguish between microphylls and megaphylls.
14. Distinguish between the homosporous and heterosporous condition.
15. Explain why seedless vascular plants are most commonly found in damp habitats.
16. Name the two clades of living seedless vascular plants.
17. Explain how vascular plants differ from bryophytes.
18. Distinguish between giant and small lycophytes.
19. Explain why whisk ferns are no longer considered to be “living fossils.”
20. Describe the production and dispersal of fern spores.
Student Misconceptions
21. Many students have difficulty in understanding the significance of derived characters that are shared between two extant groups. Just as many members of the general public have the mistaken notion that humans evolved from chimpanzees, some students will think that charophyceans are in some sense ancestral to plants or that charophyceans are identical to the last common ancestor that plants and charophyceans shared.
22. It is important to make sure that your students understand alternation of generations in bryophytes and seedless vascular plants. Plant life cycles are challenging for all students. Without a good understanding of the life cycles of plants with recognizable gametophytes and sporophytes, students will have great difficulty with gymnosperm and angiosperm life cycles.
23. Students tend to think of derived traits as “advanced.” Be careful to avoid this term. Point out that organisms have a combination of primitive and derived traits, and that all living organisms have an equally long evolutionary history, dating back to the origin of life on Earth.
24. Many students are not very familiar with or knowledgeable about plants. Some of the terminology of plant life cycles can be confusing to such students. Clarify for students the meaning of these pairs of terms:
a. homosporous and heterosporous
b. bryophyte and phylum Bryophyta
c. rhizoid and root
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