PHOTOSYNTHESIS

Introduction:
o Pigments are "molecules that absorb specific wavelengths (energies) of light and reflect all
others."
o A photosynthetic pigment (accessory pigment; chloroplast pigment; antenna pigment) is a
pigment that is present in chloroplasts or photosynthetic bacteria and captures the light energy
necessary for photosynthesis. They function by function by extending the range of light
wavelengths that a plant absorbs.
o Paper chromatography is a technique used to separate a mixture into its component molecules.
The molecules migrate, or move up the paper, at different rates because of differences in
solubility, molecular mass, and hydrogen bonding with the paper.

Objectives:
o To be able to extract the different pigments from the Papaya leaves

Types of pigments found in a leaf:

o Carotene (yellow)
o Phaeophytin (yellow-gray)
o Xanthophylls (yellow-brown)
o Chlorophyll B (green)
o Chlorophyll A (blue green)

Materials
o 1:1 petroleum ether acetone
o Strip of Whatman #1
o Papaya leaves
o Masking tape/rubber stopper with paper clip
o 1 Test tube
o Mortar and pestle
o Acetone

Methods: (Lihog lang ko ipa flow diagram thanks)

o Weigh 10g of finely cut Papaya leaves
o Crush the leaves using a mortar and pestle while adding 3mL of acetone
o Use a capillary tube and proceed on spotting on a strip of Whatman #1 paper
o Place the Whatman #1 inside a test tube filled with 1:1 petroleum ether- acetone and covered with
a stopper or tape.
o Let the chromatographic process take place
o Remove from tube if solvent front is 1 inch away from the top

Discussion:
o In paper chromatography the pigments are dissolved in a solvent that carries them up the paper.
To separate the pigments of the chloroplasts, you must use an organic solvent. Therefore in this
experiment we used 1:1 petroleum ether acetone.
o Acetone breaks down chlorophyll's lipid bonds to a plant's thylakoid structure and suspends the
pigment in solution.
o Pigments and the solvent system travels the filter paper by capillary action.
o Only one color was present due to method and systematic errors.
 o Even if one repeats the chromatographic separations, each for a different length of time, the
pigments would migrate a different distance on each run. However, the migration of each pigment
relative to the migration of the solvent would not change. This migration of pigment relative to
migration of solvent is expressed as a constant, Rf (Reference front). It can be calculated by using
the formula:

=

Solvent Front 48 mm
Distance travelled by the green pigment 32 mm
Rf 0.67mm

And in this experiment:

32
=
48

= 0.67

Conclusion:
o The experiment showed little to no separation of pigments in which the only color seen was
green. This might be a cause of a method error or instrumental and systematic errors
encountered. For example, the contamination of the used solutions, the absence of a rubber
stopper, etc.