|
Chromatography
of Simulated Plant Pigments |

Introduction
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.