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Lab Report Chemist 3

This document describes a plant pigment chromatography experiment. It introduces how plants perform photosynthesis to convert sunlight into chemical energy stored as glucose. Chlorophyll and other pigments like carotenoids and anthocyanins absorb specific light wavelengths during photosynthesis. The experiment uses paper chromatography to separate spinach pigments based on solubility. Absorption spectra of isolated pigments are then measured to identify them.

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Aiman Athirah Binti Hasbullah E21A0446
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39 views3 pages

Lab Report Chemist 3

This document describes a plant pigment chromatography experiment. It introduces how plants perform photosynthesis to convert sunlight into chemical energy stored as glucose. Chlorophyll and other pigments like carotenoids and anthocyanins absorb specific light wavelengths during photosynthesis. The experiment uses paper chromatography to separate spinach pigments based on solubility. Absorption spectra of isolated pigments are then measured to identify them.

Uploaded by

Aiman Athirah Binti Hasbullah E21A0446
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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Plant Pigment Chromatography

INTRODUCTION
As primary producers in the food chain with some bacteria and algae, plants
produce their own food by using the sun’s energy to transform carbon dioxide and
water into glucose. In this process of photosynthesis, plants convert the sun’s
energy into chemical energy that is stored in the bonds of the glucose molecule.
This energy fuels the metabolic processes of cells and is essential for life on earth.
Glucose is a simple carbohydrate that provides immediate fuel to cells but it is also
a building block for more complex carbohydrates stored by living organisms for
future use.

For photosynthesis to transform light energy from the sun into chemical energy
(bond energy) in plants, the pigment molecules absorb light to power the chemical
reactions. Plant pigments are macromolecules produced by the plant, and these
pigments absorb specified wavelengths of visible light to provide the energy
required for photosynthesis. (Appendix A) Chlorophyll is necessary for
photosynthesis, but accessory pigments collect and transfer energy to chlorophyll.
Although pigments absorb light, the wavelengths of light that are not absorbed by
the plant pigments are reflected back to the eye. The reflected wavelengths are
the colors we see in observing the plant. (Example: green pigments reflect green
light) Plants contain different pigments, and some of the pigments observed
include:

chlorophylls (greens)
carotenoids (yellow, orange red)
anthocyanins (red to blue, depending on pH)
betalains (red or yellow)

The process of chromatography separates molecules because of the different


solubilities of the molecules in a selected solvent. In paper chromatography,
paper marked with an unknown, such as plant extract, is placed in a developing
chamber with a specified solvent. The solvent carries the dissolved pigments as it
moves up the paper. The pigments are carried at different rates because they are
not equally soluble. A pigment that is the most soluble will travel the greatest
distance and a pigment that is less soluble will move a shorter distance.
The distance the pigment travels is unique for that pigment in set conditions and is
used to identify the pigment. The ratio is the Rf (retention factor) value. Standards
are available for comparison. (Appendix B)
distance pigment travels (cm)
Rf = distance solvent travels (cm)
The bands derived in paper chromatography contain the pigments found in the
plant. The bands can be cut apart, and placed in alcohol to elute the pigment in an
extract. Each pigment can be tested to derive the wavelength absorption
spectrum for that pigment. A spectrophotometer measures the absorption of light
by an extract containing the pigment and provides information that is plotted in a
graph to illustrate the absorption spectrum for the isolated pigment.

EQUIPMENT AND MATERIALS

 2 or 3 fresh spinach leaves


 Ruler
 Large test tube
 Cork with push pin
 Chromatography paper (precut 18 cm strips)
 Pencil
 Copper penny coin
 Chromatography solvent (9:1 petroleum ether & acetone)
 6 ml syringe
 Colored pencils
 Calculator
 Scissors
 Plastic wrap
 70 % Isopropyl alcohol
 Plastic pipettes
 5 test tubes (20-30 mL)
 Test tube rack
 Sharpie markers or tape (for labeling test tubes)
 4- 6 spectrophotometer cuvettes
 Test tube rack for cuvettes
 Kimwipes
 Genesys 20 Spectrophotometer

SAFETY
Wear goggles and aprons when working with chemicals.
Petroleum ether, acetone and alcohol are volatile and flammable.
Avoid breathing vapors of the reagents.

Day One
Work in teams of two for this activity. Make sure the work area is clean and dry

Preparation of the Sample:

1. Take a strip of chromatography paper approximately 18 cm long. One end


is blunt and the other is pointed.
2. With a pencil lightly make a line 2 cm from the pointed end of the paper.
3. Bend the strip of paper at the blunt end and attach it to the small end of the
cork with the push pin. Adjust the length of the paper so that when it is
inserted into the test tube, it will touch the bottom without curling.
4. Place a ruler over the leaf so that is covers the pencil line on either end.
5. Using a penny coin, press down firmly and roll along the ruler edge several
times to form a definite green line.
6. Allow the green line to dry thoroughly.
7. Use a fresh area of the leaf and repeat several times until the pencil line is
covered completely with a narrow green band. Be careful not to smear this
green line.

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