TESTING FOR BIOLOGICAL MOLECULES

AQEELAH ISAACS

3942341

BTN223

DUE DATE: 14 AUGUST 2020

 CHEMICAL TESTS FOR BIOLOGICAL MOLECULES

ABSTRACT

This experiment was done to test for the presence of biomolecules in different food samples by
using chemical indicators for each carbohydrate, protein, lipid, and nucleic acid. The results
would show a particular change of color in the biomolecule. The positive sample for that
biomolecule will be confirmed by a color change.

Starch was tested using an iodine reagent. The hypothesis was that the starch solution would
change blue/black and the rest would remain yellow. Results tested positive for starch in starch
sample and negative for the rest.

Protein was tested with a Biuret reagent. It was hypothesized that albumin would turn
purple/violet color and the others remain light blue. Results concluded that albumin was the
only one that tested positive for proteins.

For testing lipids, the reagent ethanol was used. The hypothesis was that only the cooking oil
would show a color change. It was verified in the results and cooking oil was the only sample
containing lipids.

In testing for sugars Benedict’s reagent was used. It was hypothesized that fruit juice and
sucrose would turn a yellow/orange color. To conclude the results proved the hypothesis to be
true. Fruit juice and sucrose were the only samples containing sugars.

INTRODUCTION

Biomolecules are compounds formed by living organisms and by cells. They have a broad
variety of sizes, shapes and functions. Carbohydrates, proteins, lipids, and nucleic acids are the
four primary categories of biomolecules (Rogers. K, 2020).

Carbohydrates are made from carbon, hydrogen, and oxygen. They give energy and cell support
in plant cells, as well. Carbohydrates are classified into three groups depending on the number
of sugar molecules: monosaccharides (glucose, fructose, galactose), disaccharides (sucrose,
lactose, maltose), and polysaccharides (starch, glycogen). Monosaccharides are simple sugars
containing only one molecule. Disaccharides contain two monosaccharides and polysaccharides
contain three or more monosaccharides. Benedicts test is used to find sugars.

Proteins are built with monomer units called amino acids. Amino acids are covalently bonded in
chains to form polymers of amino acids. There are 20 different amino acids that can make up a
protein (Berg. JM, 2002). The method of testing for protein is doing a biuret test.

Lipids are located in the membranes of cells, and are a store of energy. They are often
hydrophobic or insoluble in water. The lipids are composed of oxygen, hydrogen and carbon.
When there are only single bonds in the hydrocarbon chain between adjacent carbons, it is said
that a fatty acid is saturated. The fatty acid is said to be unsaturated when the hydrocarbon
chain has a double bond. They consist of a fats and oils, waxes, phospholipids, steroids. (Raven.
P, 2014). Ethanol is used for a lipid test.

METHODS AND MATERIALS

Each of the 5 test tube samples were labelled with starch, albumin, cooking oil, fruit juice and
sucrose.
a) Lugol’s/Iodine test for starch
Each test tube was filled individually with a different sample. Using a pipette, 3-5 drops
of Iodine were added to each of the different samples in the test tube. The color of the
samples were observed and recorded.
b) Biuret test for protein
Each of the 5 samples were added to an individual test tube. Using a pipette, 3-5 drops
of biuret solution were added to each of the different samples in the test tube. The
color of the samples were observed and recorded.
c) Ethanol test for lipids
Each of the test tubes were filled with one of the samples. 2cm3 of ethanol was
measured and poured into each different sample. Each Sample were shaken vigorously.
2cm3 of water was measured and poured into each test tube. Each of the samples were
observed and results were recorded.
 d) Benedicts test for reducing sugars
Using a pipette, 3-5 drops of benedict’s solution were added to each test tube
containing the different samples. A water bath was prepared in a beaker. Each of the
samples were added into the water bath simultaneously to boil for 5 minutes. Test
tubes were cooled and the color of the samples were observed and recorded.
e) Benedict’s test for non-reducing sugars
2cm3 of each sample solution were added to the test tubes. 1cm3 of dilute hydrochloric
acid was added and boiled for 1 minute. The tubes were left to cool and sodium
carbonate solution was added for neutralization. 3-5 drops benedict’s solution were
added to each test tube and left to boil for another 5 minutes. Color change was
observed and recorded.

RESULTS

The results in table 1. Showed that starch solution was the only sample that turned yellow then
a blue/black color showing a presence of starch, while the other samples remained the
yellow/orange color of iodine.

The biuret test in table 2. indicates that albumin had a color change to blue and a final color of
purple/violet showing the presence of protein. The rest of the samples remained the color blue
of the biuret solution.

Ethanol test in table 3. All of the samples besides cooking oil had no change. The test tube
containing cooking oil resulted in a white emulsion forming, indicating lipid presence.

The fruit juice in table 4. changed to a yellow/orange color after boiling, showing a positive
result for reducing sugars and the rest of the samples turned to the color of benedict’s solution.

The second Benedict ’s test in table 5. showed a positive result for non-reducing sugars only in
the sucrose solution as the color changed to yellow/orange after Benedict’s reagent was added
and boiled.
 Sample tested Iodine test
Observation Conclusion
Starch Color change to blue/black Starch present
Albumin/egg white Color change yellow No starch present
Cooking oil Color change yellow No starch present
Fruit juice Color change yellow No starch present
Sucrose Color change yellow No starch present
Table 1. Iodine test for the presence of starch.

Sample tested Biuret test

Observation Conclusion
Starch Color change light blue No protein present
Albumin/egg white Color change purple/violet Protein present
Cooking oil Color change light blue No protein present
Fruit juice Color change light blue No protein present
Sucrose Color change light blue No protein present
Table 2. Biuret test for the presence of proteins.

Sample tested Ethanol test

Observation Conclusion
Starch No color change No lipids present
Albumin/egg white No color change No lipids present
Cooking oil White emulsion formed Lipids present
Fruit juice No color change No lipids present
Sucrose No color change No lipids present
Table 3. Ethanol test for the presence of lipids.

Sample tested Benedicts test

Observation Conclusion
Starch Color change light blue No sugars present
Albumin/egg white Color change light blue No sugars present
Cooking oil Color change light blue No sugars present
Fruit juice Color change yellow/orange Sugars present
Sucrose Color change light blue No sugars present
Table 4. Benedict’s test for the presence of reducing sugar.

Sample tested Benedicts test

Observation Conclusion
Starch Color change light blue No sugars present
Albumin/egg white Color change light blue No sugars present
Cooking oil Color change light blue No sugars present
Fruit juice Color change light blue No sugars present
Sucrose Color change yellow/orange Sugars present
Table 5. Benedict’s test for non-reducing sugar.
DISCUSSION
In the test for starch, iodine will react with the starch to form a blue/black color. The amylose in
starch is responsible for that color. This reaction is the product of polyiodide chains formed
from the starch and iodine reactions.

Proteins contain peptide bonds that link amino acids together. When we tested for protein,
albumin obtained a positive result with a purple color. The other samples did not turn purple
because it needed peptide bonds to cause the reaction. The color is due to the copper ions in
the biuret solution reacting with the peptide bonds in the protein of albumin. Copper II was
able to form a complex in the alkaline solution with the peptide bonds. Once the complex was
formed, the solution turned from blue to purple.

In the lipids test cooking oil was the only substance that formed a white emulsion, whereas the
other samples had no change. The explanation being that lipids are soluble in organic solvents
and ethanol is an organic substance. Proteins, carbohydrates and nucleic acids are negative for
lipids as they are insoluble in ethanol therefore did not cause any reaction to for the white
emulsion.

During the Benedict’s test, when the sample of fruit juice was boiled in dilute hydrochloric acid,
the disaccharides are hydrolyzed to their constituent monosaccharides. The hydrolysis
monosaccharide products reduced sugars and reduced the copper in the presence of alkali
causing the change in color to yellow/orange.

CONCLUSION

It can be concluded that the results in the experiment is according to hypothesis. The starch
solution contained starch, albumin contained protein, cooking oil contained lipids, fruit juice
contained reducing sugar and sucrose was a non-reducing sugar. There was a change in color
for each biomolecule specific for their chemical reagent.
REFERENCES
 Rogers, K. (2020). Biomelecule. Encyclopedia Britannica. [Online]. Available at:
https://www.britannica.com/science/biomolecule (Accessed 13 August 2020)
 Berg JM, Tymoczko JL, Stryer L. Biochemistry. 5th edition. New York: W H Freeman;
2002. Chapter 3, Protein Structure and Function. Available from:
https://www.ncbi.nlm.nih.gov/books/NBK21177/
 Raven, P. H., Johnson, G. B., Mason, K. A., Losos, J. B., and Singer, S. R. (2014). The
chemical building blocks of life. In Biology (10th ed., AP ed., pp. 32-58). New York, NY:
McGraw-Hill.