NAME: Daniela Velez Jimenez

DATE: 09/21/2023
Laboratory Report 3 (Qualitative tests for biological molecules)
Procedure 3.1 Benedict’s test for Reducing sugars

Tube #11

Image 1. Results for Benedict’s test.

Procedure 3.2 Iodine test for Starch

Image 2. Results for Iodine test.

Procedure 3.3 Biuret test for Proteins

Image 3. Results for Biuret test.

Procedure 3.4 Lipid test

Image 4. Results for Lipid test.

QUESTIONS IN COMMON FOR ALL PROCEDURES:
1. Which tubes are the positive and negative controls?
For each procedure, tube #1 is the negative control, and tube #2 is the positive control:

 Reagents Procedure 3.1: Tube #1= Distilled water (dH2O); Tube #2= Known reducing
sugar solution
 Reagents Procedure 3.2: Tube #1= Distilled water (dH2O); Tube #2=Known starch
solution
 Reagents Procedure 3.3: Tube #1= Distilled water (dH2O); Tube #2=Known protein
solution (Casein)
 Reagents Procedure 3.4: Tube #1= Distilled water (dH2O); Tube #2=Known lipid
solution (Vegetable oil)

2. What is the difference between a result and an interpretation?

During this laboratory, the result is the reaction obtained after the mixture of the reagents, in
other words, is the observable change of the solutions (either a change of color or the formation
of layers). On the other hand, the interpretation is the meaning of this result, that is, what these
changes tell about the composition of each substance.
3. Why use dH20 as the negative control?
Distilled water is free of any contaminants and will not react with the indicator reagents because
it does not contain the substances being tested, for that reason, it is perfect to ensure that the
result is the absence of the macromolecules tested.
QUESTIONS ONLY FOR PROCEDURE 3.1:
1. Which test tubes contain reducing sugars?
Test tubes #2, #4, and #11 contain reducing sugars.
2. What purpose does the boiling water bath serve?
Heating is necessary to provide the energy needed for the reaction between Benedict’s reagent
and the reducing sugar to occur. Therefore, boiling water bathing ensures that the temperature
is at a constant level throughout the experiment, helping to prevent overheating.
 3. Why should the test tubes be in the boiling water bath at the same time?
The boiling water bath also ensures that all tubes are heated uniformly, but if the samples are
not put at the same time this property would be lost and it would give inaccurate results.
4. Would heating the contents of a cell (or organelle) be an appropriate way to bring
about or increase the rate of a biochemical reaction? Instead of an increase in
temperature, what substances do cells use to achieve the same result?
Yes, heating the contents of a cell increases the kinetic energy of the molecules, that is, the
molecules start moving faster increasing, at the same time, the rate of the reaction. However, it
is important to know that some molecules are more sensitive to heat than others; for example,
proteins can lose function and cause irreversible damage to the cell, while lipids and
carbohydrates can resist higher temperatures for longer time. In another hand, some cells use
enzymes to increase the rate of the reaction, these are proteins that reduce the activation
energy required for the reaction to occur.
QUESTIONS ONLY FOR PROCEDURE 3.2:
1. Which tubes contain starch?
Tubes #2, and #7 contain starch.
2. Why do some of the positive results seem to have only a slight purple color?
Because the starch solution must have been very well shaken, because starch granules can
precipitate, and the concentration of starch in the sample is not strong enough for a dark purple
color. In our group’s results, if tube #7 is shaken, the color of the result turns darker, because
the iodine molecules can react with the starch molecules.
QUESTIONS ONLY FOR PROCEDURE 3.3:
1. Which test tubes contain protein?
Tubes #2, and #6 contain protein.
2. Are peptide bonds found in free amino acids? How can you tell?
No, peptide bonds are not found in free amino acids, therefore, tube #3 (amino acid solution) did
not produce a positive result in the Biuret test, that is, a purple color. Instead, it stayed blue,
which means that there are no peptide bonds that bind with the copper ions in the Biuret
reagent.
QUESTIONS ONLY FOR PROCEDURE 3.4:
1. Which tubes contain lipids?
Tubes #2, and #5 contain lipids.
2. Why are the words miscible and immiscible used in this procedure?
These terms are used because in this procedure two liquid substances are being mixed.
Miscible refers to the ability of liquids to dissolve in each other and form a homogenous mixture,
while immiscible refers to the opposite, which results in a separation of liquids forming a layer.
QUESTIONS ABOUT UNKNOWN SUBSTANCES:
1. Why is it necessary to thoroughly wash all the glassware before beginning this
exercise?
To prevent the interference of contaminants in the samples, which can lead to the identification
of an incorrect substance.
2. What is the most logical choice for a negative control? Why? And why is it
necessary to incorporate negative controls in this exercise?
Distilled water is the most logical choice for a negative control, because is free of any
contaminants and will not react with the indicator reagents because it does not contain the
substances being tested, for that reason, it is perfect to ensure that the result is the absence of
the macromolecules tested.
It is necessary to incorporate negative controls in this exercise because it ensures that the
indicators are working properly so the rest of mixes are being carried out using a correct
indicator, verifying the validity of the tests and the obtained results.
3. After several previous trials, the tubes with positive results will be designated the
positive control. What is the most logical choice for positive control? Why? And
why is it necessary to incorporate positive controls in this exercise?
The most logical choice for positive control in the identification of macromolecules is a solution
that contains high concentrations (and a known amount) of the macromolecule being tested,
that way the scientist can be sure and is able to identify the presence of it in any substance.
Positive controls are necessary in this exercise to ensure that the test is sensitive enough to
detect the desired macromolecules; it also ensures that the indicators are working properly, as
well as the conditions in which the test is being carried out.

4. What is the total volume in each of the tubes? Why is this important?

 Benedict’s test: 1ml of reagent + 3ml of Benedict’s reagent
 Iodine test: 1ml of reagent + 5 drops of Iodine reagent
 Biuret test: 3ml of reagent + 1ml of Biuret reagent
 Lipid test: 3ml of reagent + 3ml of distilled water
The volume is important because it affects the sensitivity of each test. The concentration of
each macromolecule present in the samples can influence the color change produced by each
reaction, for that reason is important to use the same volume of test solution in all tubes, in
other words, it would ensure the accuracy and comparability of the results.

5. What group(s) obtained positive results for sample A? Sample B? What

conclusions can be drawn from these results? What conclusions cannot be drawn
from these results?
 Results and conclusions:
  Sample A: Obtained positive result for Benedict’s test, which means it contains reducing
sugars, and does not contain starch, proteins or lipids.
 Sample B: Obtained negative results for all the tests, which means it does not contain
reducing sugars, starch, proteins, or lipids.
From these results we cannot conclude which are the exact reducing sugars contained in
sample A, and we cannot conclude what are the exact substances evaluated.