Photosynthesis

Photosynthesis
Franclene Milla, Chanel Mirafuentes, Gianica Monteagudo, Bradley Ong
BIO 21, LAB1A MHAB1
Submitted to: Professor Elena Ragragio
Submitted: September 20, 2012

ABSTRACT
Photosynthesis is a complex process that
autotrophs or food-producing organisms
undergo by combining carbon dioxide
with water to reduce it to form glucose, a
molecule that stores energy[2]. The
various factors that are needed for such a
process and the different roles that they
play have been witnessed in this
experiment. Three parts of the procedure
were concentrated on chlorophyll as a
driver, light as an energy source and
carbon dioxide as raw material. Several
pigments present in the chloroplasts of
leaves were identified as absorbers and
channels of energy. It was also affirmed
that oxygen was a by-product of
photosynthesis.
1. Introduction
As with any living organism, plants need
energy to perform the different processes
that keep it healthy and functional. One
source is glucose, a molecule containing
considerable amounts of energy. Plants can
obtain glucose easily because they are
capable of manufacturing in a process called
photosynthesis. Photosynthesis is a very
complicated procedure with numerous subprocesses yet it cannot occur without several
factors, namely chlorophyll, light, water, and
carbon and dioxide. These factors will serve
as the foci of this activity.
This experiment aims to:
(1) Determine the role of chlorophyll,
light and carbon dioxide in
photosynthesis.
(2) Observe oxygen liberation during
photosynthesis.

(3) Demonstrate how chloroplast

pigments are separated and to
identify these pigments.
If a plant is sufficiently exposed to light,
chlorophyll is present in the leaves and there
is ample supply of carbon dioxide, the plant
is able to undergo photosynthesis.

2. Methodology
2.1. The Role of Chlorophyll in
Photosynthesis
We boiled the leaf for 10 minutes. The
boiled leaf was immersed in a test tube in
with 95% ethyl alcohol. We placed the test
tube in a hot water bath until the pigments
were extracted. The bleached leaf was
washed with water and tested for presence
of starch using IKI solution.
2.2. The Role of Light in
Photosynthesis
We place a potted plant in the dark for 48
hours. After this, we selected several leaves
and wrapped portions of them with black or
carbon paper fixed by paper clips. Then the
whole plant was exposed to light for about 5
hours. The leaf was plucked and tested for
starch using IKI solution.
2.3.The Role of CO2 in Photosynthesis
We labeled three test tubes A, B and C and
filled these with previously boiled then
cooled distilled or tap water. We put
Hydrilla sprigs into test tubes A and B in an
inverted position. A pinch of NAHCO3 was
added in test tubes B and C. We exposed the
3 test tubes to bright light. We determined if
bubble evolution occured in test tubes And
B. The Hydrilla sprigs in test tubes A and B

Photosynthesis
were removed. Five drops of phenol red
were added to each test tube (A, B & C) and
shake gently. (Phenol red changes to
yellow at low pH). We compared the color
intensities of the solutions in the test tubes.
2.4. Oxygen Liberation in Photosynthesis
We filled a one-liter beaker with tap
water. To enrich CO2 content of water, we
blew air gently into the beaker for 3-5
minutes using a plastic straw. We cut the
ends of Hydrilla under water and insertde a
thistle funnel over the Hydrilla sprigs. The
cut ends of the plant should face the tube of
the funnel. We placed a test tube filled with
water upside down over the tube of the
funnel partially dipped in thewater of the
beaker. We observed for bubbles emerging
out from the cut ends.
2.5. Separation of Chloroplast Pigments
To prepare the leaf extract, we got 15-20
mature Hibiscus leaves and cut into small
pieces. We put them in a mortar with a
pinch or two of sand. The extract was
expressed. We put in a vial and set aside.
To prepare the filter paper strip, we fit a
piece of paper into a clean, dry test tube.
With this paper as a pattern or mold, we cut
the filter paper strip. We held the filter paper
by the edges. Refrain from touching the
filter paper. (Fingerprints on the surface of
the paper may affect the chromatogram.)
The filter paper should easily slide in and
out of the tube without touching the sides.
With a pencil, not a pen, we marked about 2
cm from the top and 1 cm from the base of
the strip. We cut a point on the base of the
strip. We hung the strip on a cork by means
of a hook made from a paper clip. We
marked the outside of the test tube to show
proper solvent level. We removed the paper
strip from the dry test tube and lay it on the
table. In the meantime, we poured the
solvent into the dry tube up to the marked
line. We replaced the cork (without the
paper strip) and kept inside.

To prepare the chromatogram, we dipped a

capillary tube or a fine-tip dropper into the
vial containing the extract. We placed the
drop that comes out of the capillary tube
onto the center of the 1 cm mark at the base
of the strip. Let it dry completely by waving
the paper in the air. We added another drop
directly on the first. The green spot should
be as small as possible. We dried and
repeated the process about 10-15 times until
the spot became saturated with the green
extract. We took note that the spot must be
completely dry before each drop was added.
When we were finished, the paper had a
small, dark green spot in the center of the 1
cm mark. The spot did not reach the sides of
the strip.
When the spot wass completely dry, we
removed the cork from the tube and placed
the paper strip on the hook and inserted into
the test tube containing the solvent mixture
95 parts petroleum ether and 5 parts acetone.
The solvent level was below the 1 cm mark.
We replaced the cork with the strip and
observed the flow of the solvent until the 2
cm mark. Immediately, we removed the
paper from the tube and let it dry.
When the strip was completely dry, the
chromatogram showed the different
pigments present in the specimen.

3. Results
3.1. The Role of Chlorophyll in
Photosynthesis

Photosynthesis
The green pigment extracted was
chlorophyll. Starch was detected in the
green areas while there were only
minimal amounts in the white areas.
3.2. The Role of Light in Photosynthesis

The leaf that had been covered was

almost or completely devoid of starch.
The test showed that the leaf only
contained minimal amounts of starch.
3.3. The Role of Carbon Dioxide in
Photosynthesis
Bubble evolution occurred in test tubes A
and B. However, the rate at which said
bubbles were released was slow and had
long intervals. The table below shows the
color of the solutions after phenol red has
been added.
Table 1. Color Intensities of Solutions in A, B, and C

Test Tube Label

Color

Test Tube A

Light orange

Test Tube B

Magenta

Test Tube C

Magenta

3.4. Oxygen Liberation in Photosynthesis

Bubble evolution occurred at an extremely
slow rate with an average of 1-2 bubbles
released per minute.
3.5. Separation of Chloroplast Pigments
Table 2. Pigments Present in Hibiscus

Color

Pigment

Green-Blue Green

Chlorophyll A

Yellow Green-Light
Green

Chlorophyll B

Orange

Carotene

Yellow

Xantophyll

4. Discussion

4.1. The Role of Chlorophyll in

Photosynthesis
The leaf was boiled prior to extraction to
cause the chloroplasts to burst thus exposing
the pigments. The green areas mentioned
earlier were the photosynthetic areas
because they contain chlorophyll, an
important component of photosynthesis.
Starch test was employed because the
presence of starch indicates the occurrence
of photosynthesis. Starch was evident in the
green areas because they contain abundant
amounts of chlorophyll and therefore are
able to undergo photosynthesis. Meanwhile,
the white areas have only limited reserves of
chlorophyll. In conclusion, only areas with
chlorophyll can undergo photosynthesis.

4.2. The Role of Light in

Photosynthesis
The plant was placed in the dark to consume
its starch reserves. Some leaves were also
covered to prevent light penetration. The
uncovered portions of the leaves responded
positive to the starch test because these are
the only parts that were able to absorb
enough light to trigger photosynthesis. The
covered portions, having used up their starch
reserves for energy consumption and having
been unable to capture light energy were
ultimately unable to begin food production.
Therefore, light is needed in photosynthesis.
4.3. The Role of Carbon Dioxide in
Photosynthesis

The bubble formation that occurred in

test tubes A and B was actually the
release of oxygen gas as the Hydrilla
sprigs underwent photosynthesis. The
addition of phenol red was to identify
the pH of the solutions while
photosynthesis was occurring. Solution
A (from test tube A) turned light orange
which signifies a low pH or it was
acidic. Carbon dioxide which forms
carbonic acid with water wasnt being
consumed at the same rate as in the other

Photosynthesis
set-up. On the other hand, solution B
was magenta which means that it was
considerably more basic. The addition of
sodium bicarbonate to the solution
increased the rate of photosynthesis
hence carbon dioxide was used up more
rapidly by the sprig in set-up B than in
set-up A. This led to the solution being
slightly more basic. Solution C served as
the control set-up. The less carbon
dioxide present, the more basic a
solution is.
4.4. Oxygen Liberation in Photosynthesis

Inverting the test tube over the funnel

enables the Hydrilla plant to obtain
carbon dioxide from its environment,
enabling it to undergo photosynthesis.
The bubbles were oxygen gas being
released as by-product.
4.E. Separation of Chloroplast Pigments

The pigments were identified to be

chlorophyll a and b, carotene and
xanthophyll[3]. Chlorophyll a and b are
involved in photosystems I and II.
Carotenoids absorb different
wavelengths and transfer energy to
chlorophyll a. Chlorophyll is the most
soluble, followed by xanthophyll with
carotene being the least soluble[4].
REFERENCES
1

Committee on Biology 21 Laboratory

Manual FS AY 2008-2009. (2008).
Biology 21: General Botany Laboratory
Manual.
Mauseth, J. (2009). Botany: An
Introduction to Plant Biology, 4th
edition. Sudbury, MA: Jones and
Bartlett Publishers.

Chloroplast
Pigments.
(n.d.)
Photosynthesisinfo.
Retrieved
September
19,
2012
from
http://www.photosynthesisinfo.com/chl
oroplast-pigments/
B. Lowe. (1937). Experimental
Cookery from the Chemical and
Physical Standpoint. IO: John Wiley
and Sons