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Lab Manual Exp - No.6

The document outlines an experiment on extracting plant pigments using chromatography, detailing the introduction, objectives, materials, procedures, and questions for analysis. It explains how chromatography separates pigments from leaves, specifically chlorophyll and other photosynthetic pigments. The experiment aims to identify and measure the pigments present in spinach and Coleus leaves.

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70 views2 pages

Lab Manual Exp - No.6

The document outlines an experiment on extracting plant pigments using chromatography, detailing the introduction, objectives, materials, procedures, and questions for analysis. It explains how chromatography separates pigments from leaves, specifically chlorophyll and other photosynthetic pigments. The experiment aims to identify and measure the pigments present in spinach and Coleus leaves.

Uploaded by

noirella303
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Name: Date submitted:

Date performed: Ratings:

Experiment #6
Extracting Plant Pigments through Chromatography

I. Introduction:
Chromatography is a technique used to separate and analyze mixtures by passing them
through a special material. A mixture is dissolved in a fluid called the mobile phase, which moves
through a stationary phase like paper or a solid column. Different substances move at different
speeds, causing them to separate. This method helps identify and study individual components,
such as inks, dyes, or chemicals in solutions. Chromatography of leaves is an experiment conducted
to separate the photosynthetic pigment in the leaf. A pigment is simply a molecule that absorbs and
reflects light. Photosynthetic pigments are usually present in chloroplasts and influence the colour of
leaves. Plant leaves contain several common types of photosynthetic pigments, which are
chlorophyll includes chlorophyll a (dark green) and chlorophyll b (yellowish-green), xanthophylls
(yellow) and carotenoids (orange).

II. Objectives:

● Perform chromatography to separate a mixture of pigments from plants.

III. Materials:
● Chromatography paper strips or filter paper about 1cm x 15cm in size
● Acetone (solvent)
● 150 ml beaker or a tall glass jar
● Aluminum foil to serve as cover for the beaker
● Fresh spinach leaf
● Coleus leaf or other leaf that is red in color
● Coin
● Pencil
● Ruler
● Coffee strrer or plastic stick

IV. Procedures:
1. Using a pencil, draw a base line that is 2cm from the bottom of the paper strips. Be careful in
handling the chromatography paper as oil from the human skin alter the results. List the
paper only by its sides and be careful not to touch its front.
2. Place the spinach leaf over the paper. Pressing hard, roll the edge of the coin, and rub the
leaf into the paper, following the path of the line. Repeat until the line turns very dark.
3. Repeat the same process for the Coleus leaf using a second strip of chromatography paper.
4. Add enough acetone to cover the bottom of the beaker or glass jar ( no matter than 1cm
high)
5. Attach the top of the paper strips to a pencil or a coffee stir stick. This can be done by
making a loop with the top of the paper and fastening it with a paper clip or tape.
6. Lay the pencil or stick across the top of the beaker so that it suspends the paper above the
liquid. The bottom of the paper strips must be dipped in the solvent but the solvent should
not surpass the 2cm baseline that is the point or origin.
7. Cover the beaker and allow 15-30 minutes for the solvent to rise through the strips.
8. Remove the paper strips just before the solvent reaches the top.
9. Lay the paper strips face up. Using the pencil, immediately mark the line where the solvent
stopped before it evaporates. This is called the solvent front.
10. Allow the strips to dry.
11. Before the pigments fade, mark the top of each color that you can identify.
12. Measure the distance (in mm) travelled by each pigment from the point of origin.
13. Tabulated your data. Show the following information in your table: color observed, distance
travelled, and probable pigment.

V. Table:
Color observed Distance travelled Probable pigment

VI. Questions to answer:


1. Which pigments were you able to observe in your chromatogram?

2. Why do the pigments move at different rates through the chromatogram?

3. How do the spinach leaf and Coleus leaf differ from each other in terms of their
pigments?

4. Which of the two leaves can carry out photosynthesis better? Why?

5. Why is it an advantage for plants to have different colored pigments?

VII. Generalization:

VIII. Reference:
Shama, B. K. (2007). Chromatography (5th ed.). Prakashana, Delhi.

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