Exercise 6: Primary

Products of Photosynthesis
Group 5:
Derije, Hershey
Equiza, Hazel
Genova, Erin
Genoves, Jago Jr.
OBJECTIVES

 Identifythe products produced through the

process of photosynthesis.
 Discuss some of the chemical tests used for plant
organic matter.
 Appraise
pharmacological use of chemical
compounds found in plants.
Theory

 Photosynthesis
 majorsupplier of organic compounds used in the synthesis of other
compounds (e.g. carbohydrates, proteins)
 Oxidation
 Releases stored energy to be utilized by the organism to carry out
essential metabolic processes.
 Autotrophs
 Provide the basic material and energy for all life
through photosynthesis.
Materials
 Plant leaves  Corn  Microscope
 Molasses  Peanut  Test tube holder
 Cane sugar  Cocoa  Test tube rack
 Raisin  Coconut meat  Test tubes with
 Corn syrup  Lugol’s solution stopper
 Banana  Fehling’s
 Mortar and
solution pestle
 Potato
 Concentrated
 Filter paper
 Cassava
HNO3  Scissors and
 Mongo seeds cutter
 10% NaOH
 Soya  Droppers
 Acetone
 Water bath
 Microscope
Procedures
 Carbohydrates in plants

Fehling’s Reagent
Reminders and precautions
 Make sure the opening of the test tube is away from you
or from your classmates.
 Do not boil too vigorously or do not heat over an open
flame. When using a beaker as water bath, add boiling
chips in the water bath to avoid bumping.

DISPOSAL: Dispose the solution with brick red

precipitate in the METAL RESIDUES container.
Fehling’s Reagent Test Materials

Leaf Molasses Sugar cane

Raisins
Fehling’s
Corn Syrup
Solution
 Presence of sugar in a leaf extract
Fehling’s Reagent
Cut up a leaf which has been
exposed to bright light and
crush it well with a little
water.
Record the observed color of
each tube.
Add about 3 mL of Fehling’s
solution (1.5 mL Fehling’s A
Filter the liquid into a test
and 1.5 mL of Fehling’s B)
tube about 1/3 filled.
and then boil for a minute in
a water bath
Fehling’s solution is an
indicator for the presence of
sugar.
Record results in the
BLUE  BRIGHT RED/BRICK
worksheet.
RED PRECIPITATE (AFTER
BOILING)
 Presence of sugar
 Presence of sugar in Molasses, Cane sugar, Raisin, and Corn Syrup

Fehling’s Reagent
In test tube A, put 1 mL of
molasses and 2 mL of water.
In test tube B, put a few Shake the materials well
Get 5 test tubes and label A,
crystals of cane sugar and 3 until the materials are
B, C, D, and E.
mL of water. In test tube C, thoroughly mixed.
mash a raisin and put in 3 mL
of water and in test tube D,
put 1 mL of corn syrup and 2
mL of water.

In all test tubes, add 3 mL

Fehling’s solution (1.5 mL
Observe and record your In test tube E, put 3 mL of
Fehling’s A and 1.5 mL of
results. distilled water.
Fehling’s B) and then boil for
a minute.
Reason why there is Fehling’s A and
Fehling’s B solution.

 If two solutions are mixed before, due to alkaline

nature of solution we will get a precipitate of
Cu(OH)2 and it will interfere with our test,
indicating sugar where none existing.
Reason why there is Fehling’s A and
Fehling’s B solution.
 REAGENT MATERIALS
 FEHLING'S A is a blue aqueous solution of copper(II) sulfate pentahydrate
crystals
 FEHLING'S B is a clear solution of aqueous potassium sodium tartrate (also
known as Rochelle salt) and a strong alkali (commonly sodium hydroxide).
 Equal volumes of the two mixtures are mixed together to get the
 FINAL FEHLING'S SOLUTION (deep blue colour)
In this final mixture, aqueous tartrate ions from the dissolved Rochelle salt chelate to
Cu2+ (aq) ions from the dissolved copper sulfate crystals, as bidentate ligands giving
the bistartratocuprate(II)complex. The deep blue ingredient is the bis(tartrate)
complex of Cu2+. The tartrate tetra-anions serve as a chelating agent in the solution.
Fehling’s reaction mechanism
 Carbohydrates in plants

Lugol’s Solution
Reminders and precautions
 Be cautious in cutting using a razor blade. Hold the sharp
edge using the cover. A scalpel blade with handle may be
used.

DISPOSAL: Immerse the slides containing Lugol’s solution in a

diluted solution of bleach (1:10). When the yellow or brown
color is decolorized, dispose the solution in the sink
accompanied by copious amount of water.
Lugol’s Solution Materials

Potato
Cassava
Banana

Kamote Microscope
Lugol’s Solution
 Presence of sugar in Molasses, Cane sugar, Raisin, and Corn Syrup

Lugol’s Solution
Make thin sections of the Mount in water in a slide and
following plant materials: examine with a low-power Draw and compare the starch
Banana, potato, cassava, and objective (LPO) under the grains.
kamote. microscope.

Watch the solution as it Place a drop of Lugol’s

works under and note the solution on the slide at the
Record your results.
effect as it comes contact edge of the cover glass.
with the starch grains.
Lugol’s Solution

 Described as iodine in potassium iodide.

 A solution of elemental iodine and potassium iodide in
water, named after the French physician J.G.A. Lugol.
 REAGENT MATERIALS
5 g iodine (I2 ) and 10 g potassium iodide (KI) mixed
with enough distilled water to make a brown solution
with a total volume of 100 mL and a total iodine
content of 150 mg/mL.
Lugol’s Solution

 Iodineforms a coordinate complex between

the helically coiled polysaccharide chain
and iodine centrally located within the
helix due to adsorption. The color obtained
depends upon the length of the unbranched
or linear chain available for complex
formation.
 Proteins in plants

Xanthoproteic Test
Reminders and precautions
 Concentrated nitric acid is a corrosive substance that produce
noxious gases. Avoid inhalation and contact with eyes and skin.
Wear appropriate PPE.
 Make sure that the nitric acid that will be used is colorless. Do
not use it if it is yellow (or darker) in color.
DISPOSAL:
Dispose the residue together with the filter paper in the solid wastes
bin.
Alkalinify the solution by adding a slight excess of 10% sodium
bicarbonate. Allow the fizzing (effervescence) to stop. Dilute the
water and dispose in the sink with copious amount of water.
Xanthoproteic Test Materials

Mongo beans Soya beans

Corn seeds

Litmus paper

Nitric Acid Sodium Hydroxide

 Presence of Proteins in Various Plant Materials

Xanthoproteic test

Soak a small amount of Afterwards, grind these

Add water and let stand for
mongo, soya, and corn seeds materials separately with a
30 minutes.
in water overnight. mortar and pestle.

Xanthoproteic test. To the

supernatant, add 1 mL of
concentrated nitric acid and boil
in a water bath for 5 mins.
Observe a yellow color that
appears in the solution. This
Record your results. indicates the presence of
Filter the materials.
proteins. Cool the solution and
add about 10 mL of 10% sodium
hydroxide to make it alkaline to
litmus. The yellow color deepens
to orange.
Xanthoproteic Test

 Xantho is a medical term which means “Yellow”.

REAGENT MATERIALS
Concentrated Nitric acid
10% Sodium hydroxide
Xanthoproteic Test

 Aromatic amino acids, such as Phenyl alanine, tyrosine

and tryptophan, respond to this test.
 On Nitric acid - the aromatic phenyl ring is nitrated to give
yellow colored nitro-derivatives.
 At alkaline pH, the color changes to orange due to the
ionization of the phenolic group.
 Phenylalanine gives negative or weakly positive reaction
Xanthoproteic Test reaction mechanism
 Lipids and Fatty Acids

Grease Spot Test

Reminders and precautions
 Perform this test inside a fume hood or in a
well-ventilated area far away from flammable
solvents, electrical equipment and outlets, and
smoke detectors. Do not perform without the
supervision of the laboratory instructor.
 Do not burn the paper in the procedure.
Grease Spot Test Materials
Peanuts

Filter paper
Coconut meat

Cocoa powder

Fire
Vegetable oil
 Presence of Proteins in Various Plant Materials

Grease Spot Test

Rub a peanut seed on one
corner, coconut meat on
Get a 10 cm x 10 cm filter another corner, cocoa In the center of the paper,
paper. powder on another corner. put a drop of water.
Put a drop of vegetable oil on
the other corner.

Make the grease spot test. A

grease spot often looks like a
If it is a wet spot, it will dry water spot. Hold the paper
Record your results.
out and disappear. over the flame for a few
seconds.
Grease Spot Test
 Oils and fats may be recognized by the fact that they
make a grease spot on unglazed paper.
 Most grease or fat have a high boiling point. In room
temperature, the spot of water can absorb enough heat
from the air and evaporized, unlike the spot of grease.
When the liquid is inside the sheet of paper, it diffracts
light. So, light can pass from one side of the paper to
another side. This gives the phenomenon of "translucent".
 Lipids and Fatty Acids

Chromatography
Reminders and precautions
 Acetone is a volatile solvent, which is mildly irritant. Avoid
inhalation and contact with eyes and skin. Wear appropriate
PPE.
 Place the test tube in a rack with minimal agitation. Do not
disturb while developing the chromatogram.

DISPOSAL: Air-dry the filter paper and dispose in

the trash bin. Dispose the acetone in the
NONHALOGENATED WASTS container.
Chromatography Materials

Leaf
Grass 85% Acetone

Filter paper strips

 Terpenes and Chlorophyll Pigments

Chromatography Shake the flask and allow to

stand until the color has
passed into the acetone.
When the acetone has
Chop up a handful of grass become a deep green, view
Put into a flask with about 25 the solution by reflected
cuttings or leaves of any
mL of 85% acetone. light. A characteristic blood-
tree.
red fluorescence is visible.

Prepare the chromatograph

Drop a spot of solution about
Dry and put another drop on
1 cm from the bottom of the
the same spot.
strip of filter paper.
paper by cutting a strip of
filter paper. The length of
the paper should exceed a 25
mL test tube, while the
width should be less than the
diameter of the test tube.
 Terpenes and Chlorophyll Pigments

Chromatography When the sides of the test

In the mean time, put a test
Repeat the procedure for
tube on the stand and pour 2
about 3x. Completely dry the
mL of acetone. Cover the
filter paper.
test tube with a stopper.
tube are dry, carefully insert
the strip of filter paper until
the lower edge reaches the
acetone. Make sure the level
of acetone does not reach
the drop of chlorophyll
solution.

Observe the colors that

appear as the acetone rises Allow the solution to spread Put a stopper on the test
up the filter paper strip. upward the filter paper. tube.
Record your observations.
Chromatography

 Paper chromatography is a useful technique in the

separation and identification of different plant pigments. 
 chlorophylls (greens)
 carotenoids (yellow, orange red)
 anthocyanins (red to blue, depending on pH)
 betalains (red or yellow)
 REAGENT MATERIALS
 85% Acetone
Chromatography
 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.
References
 Randy Sullivan (2012). Fehling Test. Retrieved Oct 24, 2019 from
https://chemdemos.uoregon.edu/demos/Fehling-Test
 Stack Exchange Inc. (2019). Functional difference of Benedict's solution and Fehling's solution. Retrieved
Oct 24, 2019 from
https://chemistry.stackexchange.com/questions/62034/functional-difference-of-benedicts-solution-and-f
ehlings-solution
 White Pebble Inc. (2013). Fehlings Solution. Retrieved Oct 24, 2019 from
http://gluedideas.com/content-collection/encyclopedia-americana-11/Fehlings-Solution.html
 Semanticscholar (n.d.) Lugol’s Iodine. Retrieved Oct. 24, 2019 from
https://pdfs.semanticscholar.org/7017/dbf8941c6f09f1bc1bd212f4e3165c80d408.pdf
 My Scientific Blog - Research and Articles (2014). Amino Acids. Retrieved Oct 24, 2019 from
http://upendrats.blogspot.com/2014/06/the-amino-acids.html
 Gaurab Karki (2018). Xanthoproteic test: Objective, Principle, Reagents, Procedure and Result. Retrieved
Oct 24, 2019 from
https://www.onlinebiologynotes.com/xanthoproteic-test-objective-principle-reagents-procedure-and-res
ult/
 Plant Traveling Lab (2010). Plant Pigment Chromatography. Retrieved Oct 24, 2019 from
https://www.depts.ttu.edu/ciser/science-teacher-resources/traveling-lab/curriculum/plants/Plant_Pigm
ent_Chromatography.pdf
 Biology Corner (n.d.). Separation of Plant Pigments Using Chromatography. Retrieved Oct 24, 2019 from
https://www.biologycorner.com/worksheets/plant_pigments.html