GENERAL BIOLOGY 1 (FINAL TERM)

LEARNING ACTIVITY #2 Light and Pigments

NAME: ANGELA ERANDIO SECTION: 11-STEM GAUSS

PAPER CHROMATOGRAPHY AND LEAF PIGMENTS

Ramon and Macy are good friends which regularly eats vegetables as a part of their diet. On
one occasion, Ramon’s ‘baon’ contained a spinach leaf as one of the ingredients. Both we’re
puzzled and somehow curious on why the vegetable have that deep green color compared to
other leafy vegetables. This prompted them to have the following discussion:

Ramon’s Baon of Bacon and Spinach. Ramon Looks closely at his baon and Macy was reminded of how of his
makes the following statement. vacation to japan during winter where
the trees with green leaves turned into
a different color during autumn.

Macy then made the following
statement:
Both students scoured the internet to
look for procedures on how to
determine the different pigments in
plant leaves.
Luckily, they we’re able to find a
laboratory protocol they could
follow to extract plant pigments thru
a process called Plant
Chromatography

You can access the same Laboratory module that Ramon and Macy used by clicking this link:
https://www.depts.ttu.edu/ciser/science-teacher-resources/traveling-lab/curriculum/plants/Plant_
Pigment_Chromatography.pdf
 Leaf Pigments Experiment
Primary producers in the ecosystem, such plants and phytoplankton, contain pigments that
allow them to harvest the energy from sunlight and use its energy for transforming water and
carbon dioxide into glucose and water in a process known as Photosynthesis. Plant pigments
are macromolecules produced by the plant which absorb specific wavelengths of visible light to
provide energy required for photosynthesis. The primary pigment that the plant uses is the
Chlorophyll, however, other accessory pigments are also used which absorb the energy form a
different wavelength and transfer this energy to chlorophyll. The different pigments that can be
observed in plants are
● Chlorophyll (Greens)
● Carotenoid (Yellow and Orange Red)
● Anthocyanin (Red to Blue)
● Betalain (Red or Yellow)
A process known as chromatography separates molecules based on their different solubility
on the 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 below
for comparison.

𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑝𝑖𝑔𝑚𝑒𝑛𝑡 𝑡𝑟𝑎𝑣𝑒𝑙𝑠 𝑖𝑛 𝑐𝑚

𝑅𝑓 = 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 𝑡𝑟𝑎𝑣𝑒𝑙𝑠 𝑖𝑛 𝑐𝑚

PART A
In the first part, Macy and Ramon performed a Paper Chromatography test which separates
molecules based on their different solubility on the selected solvent. Ramon and Macy’s
experiment yielded the following results:

Result A: Resulting Paper Chromatograph

Answer the Guide Questions Successively to situate which student is supported by the results
of the experiment: Always answer in COMPLETE SENTENCE.
(Recognizing Evidences)

● Besides green, what other colors were observed by Ramon and Macy, can you
enumerate them below?

- as observed by Ramon and Marsey, except for green or Chlorophylls which

are the primary pigments involved in photosynthesis and give leaves their
green color. There are also yellow and orange or carotenoids. carotenoids are
part of a group of pigments found in leaves. because During paper
chromatography, the different pigments move at different rates through the
paper based on their solubility so this separation allows for the visualization
of the various pigments present in the leaf extract.
● What do these colors tell us about the pigments present in the Leaf? How many
pigments are in the leaf based on the observed colors?

- the colors of leaves reflect the types and concentrations of pigments present,
each serving specific roles in the plant's life processe they tell us not only
contribute to the aesthetic diversity of leaves but also reflect the leaf's
adaptive strategies for light absorption, energy production, and protection
against environmental stresses. there is 2 pigments that are present that is
chlorophylls responsible for the green color and carotenoids that contribute
to yellow and orange colors

● Having read the laboratory procedures, what component in the experiment made the
colors climb up the paper and separate?

- It's the solvent that allows the colors to climb up the paper and separate,
based on the differential solubility and interactions of the components with
both the solvent and the paper.

● What can you infer about the position of Pigment 1? Is it heavier or lighter compared to
the other pigments?

- the pigment 1 and pigment 2 have similar structures but pigment 1 is slightly
smaller than pigment 2 and pigment 1 is heavier and larger than pigment 3,4,
and 5

● What can you infer about the position of Pigment 5? Is it heavier or lighter compared to
the other pigments?

- pigment 5 it is less heavy in the sense of being less retained by the paper and
more light in terms of mobility compared to the other pigments. This means
that pigment 5 has a greater affinity for the solvent and tends to move further
up the paper.
 ● Is there any way that pigment 1 can reach the height of pigment 5, briefly explain your
answer.

- The differences in polarity, size, and interactions of these pigments with the
chromatographic system make it unlikely for pigment 1 to reach the same
height as pigment 5 in the chromatography. However, adjustments in the
chromatographic conditions, such as altering the solvent composition or
using specialized techniques, might influence the separation behavior and
potentially affect the relative positions of the pigments.

(Creating Warrants in support for Claims; Creating Claims)

● What does the Rf value say about the pigments? Can different pigments have the same
Rf values?

- The Rf value provides information about the relative mobility and solubility
of pigments in chromatography. While different pigments can have the same
or similar Rf values, caution should be exercised in making definitive
identifications based only on Rf values, as other factors, such as molecular
structure and environmental conditions, can influence the
results.

● Which student’s argument do you think is supported by the results of

the Paper Chromatography? (Ramon or Macy)

- Macy's because based solely on the results of paper

chromatography, that the leaf contains chlorophyll and other
pigments is more supported .

● How can you say so? Support and Explain your answer.

- The presence of distinct bands corresponding to chlorophyll,

carotenoids, and potentially other pigments, support that leaves
contain chlorophyll and other pigments, providing a more
detailed and accurate picture of the pigments responsible for leaf
coloration.
PART B
Using the results obtained from the chromatography performed by Ramon and Macy, complete
the data table below to reveal the identity of each of the pigments observed.

Data Table 1: Description and Rf Values of Pigments

A B C D E F
Pigment Description of Color Distance Distance Rf Value Name of
Solvent Front Color (Column D / Pigment (Use
Travelled Travelled Column C) reference provided
in page 2)

5 yellow/orange 9.6 9.0 0.94 β-carotene

4 yellow 9.6 7.3 0.76 Lutein

3 yellow 9.6 5.7 0.59 Lutein

2 green 9.6 3.7 0.39 violaxanthin

1 grassy green 9.6 2.5 0.26 Chlorophyll a

(Recognizing Evidences)

● What other pigments besides Chlorophyll was Ramon and Macy able to see?

- its Carotenoids pigment that contribute to the yellow and orange colors in
photo chromatography.

● What are these other pigments responsible for?

- In chlorophyll, plants convert light energy into chemical energy to fuel their
growth and Carotenoids they help in capturing light energy for
photosynthesis protect the plant from damage caused by excessive light.
These pigments play vital roles in photosynthesis, protection from
environmental stressors, attraction of pollinators, and providing a diverse
range of colors to different plant structures
.
● Which type of pigment is the most abundant? (Relate it to the width and abundance of color
observed in the paper chromatography)

- The most abundant pigment observed in the paper chromatograph is

chlorophyll a, because the width and intensity of this green bands
corresponding to chlorophyll a would generally indicate its abundance
compared to other pigments present
-
(Creating warrants in support of Claims, Creating Claims)

● Why do you think the leaf invests energy

in producing these other pigments? How
does it help the plants in performing
photosynthesis?

- Leaves invest energy in

producing pigments beyond
chlorophyll to enhance
photosynthesis and ensure
overall survival and these
pigments serve various roles,
including photoprotection by
dissipating excess light energy, providing UV protection, attracting
pollinators and aiding reproduction.

● What if Ramon and Macy use a different type of leaf, say a Coleus blumei. Leaf pictured
below, Do you think it will also lead to the same results? Do you think chlorophyll will
still be the most abundant? (Start with showing your data, then support it with your warrants, and
lastly, state your claim answering the question.)

- I tried to find a coleus blumei plant to perform the chromatography, but I didn't find
that type of plant. but I searched to find out about this plant. If ramon and macy use
Coleus blumei in paper chromatography is expected to yield different results in
terms of pigment composition compared to other leaves. Given the genetic,
environmental, and physiological differences between plant species, it's likely that
the pigments observed in Coleus blumei. I'd assert that using Coleus blumei in paper
chromatography would likely reveal a different set of pigments compared to other
leaves. While chlorophyll might still be present and essential for photosynthesis, the
chromatogram would likely showcase a diverse range of pigments unique to Coleus
blumei, potentially altering the abundance and variety of pigments observed
compared to other plants.
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