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Bio L 2281 Experiment 6
Bio L 2281 Experiment 6
ABSTRACT
1. Describe the structures and functions of the four main categories of
biologically important macromolecules.
2. Perform chemical tests to identify the presence of lipids, proteins, two
forms of carbohydrates, and DNA.
3. Understand and explain the importance of control experiments
Experiment 6:
Biochemical Testing of Macromolecules Vy Nguyen 1
Part I: Answer the following questions. Include questions 110 in your report.
1) Present a typed version of Table 1, complete with your observations. Which of the
solutions is a negative control? What does this result tell you? (2 pts)
Table 1: Solutions and Color Reactions for Benedict’s Test and Iodine Test
The Benedict’s test is used for detecting reducing sugars, which contain aldehyde groups that are
oxidized to carboxylic acids, such as glucose, fructose, glyceraldehyde, arabinose, lactose, and
maltose. Ketoses can also be a reducing sugar if they contain alphahydroxyketones or if they
can isomerise to aldoses during the reaction. The negative control of the Benedict’s test is
distilled water
, which does not contain reducing sugars (shows what a negative result looks like)
and appears as blue. The following solutions show the negative result of the Benedict’s test:
➢ Tube C: 10 drops of 1% sucrose solution
➢ Tube E: 10 drops of 1% starch solution (control for Iodine)
➢ Tube G: 10 drops of distilled water (control)
The Iodine test is used for detecting starch. Iodine interacts with coiled polymer of glucose and
becomes black. The negative control of the Iodine test is distilled water,
which does not contain
starch (shows what a negative result looks like) and appears as yellowbrown/orange. The
following solutions show the negative result of the Iodine test:
➢ Tube A: 10 drops of onion juice
➢ Tube C: 10 drops of 1% sucrose solution
➢ Tube D: 10 drops of 1% glucose solution (control for Benedict’s)
➢ Tube F: 10 drops of milk
➢ Tube G: 10 drops of distilled water(control)
Based on both test, there is a presence of both a reducing sugar and starch in potatoes and an
absence of both a reducing sugar and starch in sucrose and distilled water. There is a presence of
Experiment 6:
Biochemical Testing of Macromolecules Vy Nguyen 2
a reducing sugar and an absence of starch in onion juice, glucose, and milk. In contrast, there is a
presence of starch and an absence of a reducing sugar in the starch solution (which is a control
for the iodine test).
2) Glucose in the urine, glycosuria, can be an indicator of diabetes. Which test could you
use to determine if a patient’s urine contained glucose? (1 pt)
The Benedict’s test is used for detecting reducing sugars, which contain aldehyde groups that are
oxidized to carboxylic acids, such as glucose, fructose, glyceraldehyde, arabinose, lactose, and
maltose. Thus, I could use the Benedict’s test to determine if a patient’s urine contains glucose.
3) How can a substance taste sweet, yet give a negative reaction with the Benedict’s test?
(1pt)
The Benedict’s test is only used for detecting reducing sugars, which contain aldehyde groups
that are oxidized to carboxylic acids, such as glucose, fructose, glyceraldehyde, arabinose,
lactose, and maltose. Sucrose is not a reducing sugar because the glycosidic bond between its
components, fructose and glucose, prevents their isomerization. While sucrose tastes sweet, it is
not a reducing sugar, and thus it will give a negative reaction with the Benedict’s test.
4) What have you learned about how carbohydrates are stored in onions and potatoes from
the experiments performed in Table 1? (1 pt)
Carbohydrates are stored as starch in potatoes and are stored as reducing sugars in onions. While
our result shows that there is a presence of reducing sugar in potatoes, the yellow color illustrates
that there is very little sugar in potatoes, in comparison to the brown color of onions. Thus
onions primarily store its carbohydrate as reducing sugars, while potatoes primarily store its
carbohydrate as starch.
5) Present a typed version of Table 2, complete with your observations. Which of the
solutions in Table 2 is a positive control? What is the purpose of this control? (list 2
purposes) (2 pts)
The protein solution (1% Bovine Serum Albumin) turns deep violet with the Biuret reagent, thus
it is the positive control. The positive control shows what a positive result should look like and
ensures that the experiment is done correctly (procedures, reagents, equipment).
6) Suppose you have tested an unknown sample with Biuret and Benedict’s reagents. The
solution mixed with Biuret reagent is blue. The solution boiled with Benedict’s reagent is
also blue. Can you conclude the identity of the sample? Why or why not? (1 pt)
No, the blue solution resulted from the Biuret test means that the solution is negative (does not
contain) for protein, and the blue solution resulted from the Benedict’s test means that the
solution is negative (does not contain) for a reducing sugar. The solution’s identity can be
sucrose, starch, or lipid.
7) What can you conclude about the molecular content of milk based on all of the tests you
performed on milk? (1 pt)
Based on all of the tests I performed on milk, the molecular content of milk includes a reducing
sugar (positive for Benedict’s Test) and protein (positive for Biuret Test). Milk does not contain
starch (negative for Iodine Test).
8) Present a typed version of Table 3, complete with your observations. What can you
conclude about the function of the detergent comparing test “N” and “O” in table 3? (In
other words, what did the detergent do?) (2 pts)
The detergent forms an emulsion, preventing water and oil to separate into two different layers.
9) Present a typed version of Table 4, complete with your observations. Could the Dische
diphenylamine test tell you if a DNA sample is contaminated with RNA? Why or why not?
(2 pts)
Experiment 6:
Biochemical Testing of Macromolecules Vy Nguyen 4
Table 4: Dische Diphenylamine Test for DNA (DEMO observed in fume hood)
4 1 ml water Clear
No, the phenylamine only reacts with the deoxyribose sugar of DNA, not the ribose sugar of
RNA, thus the Dische diphenylamine test could not tell you if a DNA sample is contaminated
with RNA.
1) Present the results of your tests in typed table format (with proper titles). Use the tables
in this handout as examples. You should describe the color of each reaction in your tables
AND indicate (+) or () for the test result. If you did not perform a test, indicate that in the
table. (2 pts)
Table 5: Identification of unknown solution
2) Write a simple summary or conclusions paragraph (45 sentences) stating the identity of
your unknown and how you ruled out other possibilities. (3 pts)
The color of the unknown solution for the Benedict’s test, which is used to detect reducing
sugars, is the same as the color for the negative control (distilled water) blue; so there is an
absence of reducing sugar in the unknown solution. The color of the unknown solution for the
Iodine test, which is used to detect starch, is the same as the color for the negative control
(distilled water) brown; so there is an absence of starch in the unknown solution. The color of
the unknown solution for the Biuret test, which is used to detect protein, is the same as the color
for the negative control (distilled water) blue; so there is an absence of protein in the unknown
solution. According to the observations of the negative control (distilled water), the unknown
solution also shows negative results for the Benedict’s Test, Iodine Test, and Biuret Test; Thus
we can rule three of the four possibilities: glucose, starch, and BSA protein. The only remaining
option, sucrose, cannot be detected using the Benedict’s Test, Iodine Test, and Biuret Test,
therefore the identity of the unknown solution is sucrose
.
Experiment 6:
Biochemical Testing of Macromolecules Vy Nguyen 6
6. What are negative and positive controls? And what are their functions?
• Positive shows that test/experiment runs correctly (variable)
• Negative shows what a negative result looks like (no variable)
Experiment 6:
Biochemical Testing of Macromolecules Vy Nguyen 7