CHEMISTRY PROJECT

Leaf Chromatography

Name : Akshit Madathil

Class : XII
 Certificate
This to certify that Akshit Madathil of standard XII has completed
the Chemistry project in partial fulfilment of the curriculum of ALL
INDIA SENIOR SECONDARY CERTIFICATE EXAMINATION (AISSCE).
This project was completed at Narayana e Techno School during
the year 2022-2023.

Internal examiner External examiner

School stamp Principal’s signature

 Acknowledgement
I would like to express my deepest regards to the principal, Mr. Ravi
Swarnkar , for providing us with the infrastructure and facilities for
the project.

I would further like to extend my thanks to my chemistry teacher

(Mr. Sachin Gorivale) under whose guidance this project was
performed. His constant support and invaluable guidance helped
me to achieve completion of this project.

I also want to express my gratitude to the Chemistry Department

of the school and Chemistry laboratory in-charge.

Last but not the least, I want to thank my family for their moral
support.
 Index
-Aim
-Introduction
-Theory
-Requirements
-Procedure
-Result
-Conclusion
-Precaution
-Bibliography
 Aim
To determine the different pigments and
compounds in a leaf using chromatography
 Introduction
Leaf chromatography is an experiment that allows us to see the
colourful pigments that leaves have hidden inside them. The green
leaf colour is the chlorophyll, which helps plants absorb the yellow
and blue wavelengths of light. Chlorophyll is the main pigment
used for photosynthesis, or harvesting energy from sunlight! Those
leaves have so much chlorophyll in them that it hides the other
pigments from view. Some of these other pigments are
carotenoids (yellow, orange, red, or brown in colour; absorb blue
light) and anthocyanin (pink-red coloured; absorb blue-green light)
– these two pigments help plants absorb additional wavelengths of
light and help plants deal with some of the stress that sunlight can
cause (Harvard Forest). In the fall, the chlorophyll in leaves breaks
down and the nutrients are reabsorbed by the plant, leaving
behind the other leaf pigments – this is why we get to see so many
gorgeous red, orange, and yellow trees! By breaking down the
leaves and separating out the pigments, we’re able to get a sneak
preview of the different colours we might see on these trees in the
fall.
 Theory
Leaf Pigments:
Plants make an amazing variety of pigment molecules, far more than
animals. After all, plants are creatures of light. They sense light to control
their growth and rapid responses to the environment, and they use light as
their source of energy. Plants produce pigments to advertise rewards for
animals which pollinate flowers and disperse seeds. Thus, pigments may
have physiological and/or biological functions.

There are three types of pigments present in the leaves of plants, and their
retention or production determines the colours of leaves before they fall from
, molecules, beyond the simple chemical formulas that describe the
numbers of atoms of different elements making up the molecule. The
example shown here is the common sugar glucose.

Glucose can be purchased as a sweetener, most commonly is one half of the

common table sugar (sucrose), which is a disaccharide. More complicated
diagrams will be displayed to illustrate the structures of the three types of
pigments that are present during the aging of leaves: chlorophylls,
carotenoids, and anthocyanin.
Carotenoids:
Carotenoids are very long-chain water-repelling pigments that are
synthesised in the plastids of plant cells. In the sunflower, a common
carotenoid, ß-carotene, is produced in the chromoplasts of the ray
flowers to produce bright yellow-orange colours. These pigments
primarily absorb in the blue wavelengths, allowing the longer
wavelengths to be scattered and producing the yellow colour. In autumn
foliage, the carotenoids are left over in the chloroplasts and revealed
from the loss of chlorophyll.


Chlorophylls:
The chlorophylls, a and b, are the pigments of photosynthesis. They are
produced in chloroplasts in the photosynthetic tissues of the leaf. The
chlorophyll molecules are very water repelling, partly because of the long
phytol tail in the molecule. The closed ring of the molecule is similar to
the haemoglobin of our blood, but holds a magnesium ion rather than
iron. It is a large and expensive molecule to make, partly because each
ring contains four nitrogen atoms. Chlorophyll is normally broken down
towards the end of the leaf life span, and much of the nitrogen is
resorbed by the plant.
Anthocyanin:
Anthocyanin are water-soluble pigments produced via the flavonoid
pathway in the cytoplasm of the coloured plant cell. The attachment of
the sugar molecule makes them particularly soluble in the sap of the
vacuole, where these molecules are stored…..once they are launched.
These are responsible for the pink-red colours of most flower petals, of
most red fruits (like apples) and almost all red leaves during the autumn.
Anthocyanin absorb light in the blue-green wavelengths, allowing the red
wavelengths to be scattered by the plant tissues to make these organs
visible to us as red.
 Requirements
- Apparatus
• Glasses / Jars / Beakers
• Mortar and pestle
• Filter paper / Coffee filter
• Plastic wrap
• Leaves (from different plants)

- Chemicals Required
• Isopropyl alcohol
 Procedure
Keep leaves from different trees separate and follow the
steps below for each set of leaves, so you can compare
results for your leaf chromatography experiment.

1. Tear up the leaves into several pieces and place them in a

beaker or glass, then add just enough rubbing alcohol to cover
them. Cover the beaker with plastic wrap to keep the alcohol
from evaporating.

2. Put the beaker in a dish of hot tap water for about 30

minutes, until the alcohol turns green as the pigments from the
leaves are absorbed into it.

3. Cut a strip of filter paper about a half inch wide and tape it to
a pencil. Suspend the pencil across the beaker and let the strip
just barely touch the alcohol and pigment mixture.
 Result
A bit of the mixture will travel slowly up the
paper.
After about 30-90 minutes you should be able
to see the “green” colour break up into several
different colours as the different pigments
begin to separate.
 Precautions
• Use gloves to prevent any possible damage of
skin
• If skin damage does occur, tend to immediate
first aid.
• Deal with chemicals with caution and avoid
contact with sensitive body parts
• Hands and utensils used should be throughly
washed after use
 Conclusion
Using chromatography we can see the different pigments
and compounds present in leaves.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.
Rf = distance pigment travels (cm) / 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.
 Bibliography
• ctsciencecentre.org
• harvardforest.fas.harvard.edu
• learning-centre.homesciencetools.com
• scienceinschool.org