Biological Macromolecules

Introduction to Biology with

Lab

Hannah Barlow

Introduction:
This experiment was meant to teach us the four kinds of macromolecules: proteins,
lipids, carbohydrates, and nucleic acids. We are to perform 4 tests which will determine
if the four macromolecules are present.
A Macromolecule is a large molecule that has small molecules that link together. There
are four kinds of macromolecules. These include: lipids, proteins, carbohydrates, and
nucleic acids.
Lipids are organic compounds that are oily when touching and are not soluble.
Examples of lipids are: waxes, oils, steroids, fatty acids. This would include fish,
sunflower seeds, canola oil, and green vegetables. Lipids are very beneficial for the
body. They allow brain function, protective layer for organs, natural steroids, and
supporting the reproductive system. In this experiment we will use Sudan III to test for
lipids on filtered paper.
Proteins are long chains of amino acids. These amino acids are compounds made up of
one amino group and one carboxyl group. There are two types: essential and
nonessential. In total, there are twenty. In this experiment, we will use Biurets reagent
to detect if there are proteins present.
Carbohydrates are the bodys main source of energy. They can be stored for quick use,
immediate use, or later use. Carbohydrates are produced in green plants during
photosynthesis. There are to types of carbohydrates: simple and complex. Simple
carbohydrates are also known as simple sugars. Complex carbohydrates are at least
three linked sugar molecules. Complex carbohydrates are often found in whole grain
foods, yams, and beans. In this experiment, we will use Benedicts reagent to find
reducing sugars that are present, and the IKI solution is used to find the starch present
in foods.
Nucleic Acids are long chains of RNA or DNA. Both RNA and DNA are called
nucleotides which include: a phosphate, sugar, and nitrogenous base. These are
essential and responsible for all the genetic makeup of all living organisms.
As the positive controls in our experiments we have: glucose for the sugar test, starch
and the potato for the starch test, vegetable oil for the lipids, and milk for protein. For
the unknown, I would say we will be able to know exactly what it is by the end of this
experiment and what category it belongs under.

Methods and Materials:

ITEM (Student Supplied)

NEEDED

Bottle of Distilled Water

Egg

1
1

Boiling Water

Roll of paper towels

Dish Soap

Pencils/ Pen

Milk, 2%

Russet Potato, raw

Oven Mitt

Fork

Plastic Spoon

ITEM (HOL Supplied)

Filter Paper, Round

12 in Aluminum Foil

Well Plate

250 mL beaker

Cleaning Brush

Test Tube, 13x100mm

Safety gloves

Safety glasses

Graduated Cylinder, 10 mL

1 plastic cup

1 sharpie

Experiment bag

Pipets

Notebook

Testing for Sugars and Starches:

1. Properly put on safety glasses and goggles.
2. Using the graduated cylinder, measure 10mL of distilled water and put it in the
bottled named UNKNOWN. Gently shake bottle to dissolve the unknown
substance into the water.
3. Separate eggs white and yolk. Catch egg whites in plastic cup.
4. Use a marker to label your six test tubes: DH2O, Albumin, Unknown, Milk,
Glucose, and Egg White Albumin stock. Place all six test tubes in the well plate
for a test tube holder.
5. Use albumin pipet to measure 0.5mL of egg whites into the test tube named egg
white albumin stock.
6. Use a pipet for albumin to measure 4.5mL of distilled water and place it into the
egg white albumin stock test tube. Swirl it around to mix it. Place the pipet used
into the Egg white albumin stock test tube.
7. Use the Albumin pipet to transfer 1mL of the egg whie albumin stock to the test
tube named Albumin.
8. Label a jumbo pipet dH2O and transfer 1mL of distilled water into the test tube.
9. Label a pipet Milk and transfer 1mL of milk into the test tube labeled milk.
10. Add 24 drops of the UNKNOWN solution to the corresponding test tube.
11. Add 24 drops of glucose to the corresponding test tube.
12. Observe the color and record.
13. Boil water. Carefully fill the 250 mL beaker full.
14. Use oven mit to protect your hands.
15. Add 40 drops of Benedicts reagent to all five test tubes.
16. Place 5 test tubes into the boiling water and allow samples to sit for 10 minutes.
17. After 10 minutes, remove test tubes from water, and return them to the well plate.
18. Observe and record the color.
19. If color is brown, a reducing sugar is present.
20. When youre done, pour all test tubes out with lots of water, and wash and dry
test tubes.
21. Use the marker to label five test tubes as: dH2O, albumin, UNKNOWN, milk, and
starch. Place all test tubes into well plate.
22. Use the Albumin pipet to measure 1 mL of the egg white albumin stock solution
into the test tube named Albumin.
23. Use the dH2O pipet to measure 1 mL of distilled water into the corresponding
test tube.
24. Use the Milk pipet to transfer 1 mL of milk into the corresponding test tube.
25. Add 24 drops of Starch from the bottle to the Starch test tube.
26. Add 24 drops of UNKNOWN from the bottle to the UNKNOWN test tube.
27. Use the spoon to get a small amount of potato. Leave the potato in the center of
the spoon.
28. Observe the color the 5 test tubes and the potato and record.

29. Use the marker to label a pipet with IKI.

30. Use the IKI pipet to measure 1 drop of IKI solution from the glass bottle into each
of the 5 test tubes and to the potato.
31. Observe and record the colors of the 5 test tubes and the potato.
32. A black color indicates the presence of starch in the test tubes. Observe and
record data.
33. When data has been recorded, pour the contents of the 5 test tubes down the
sink. Wash and dry the 5 test tubes.
Testing for Proteins and Lipids:
2. Use a marker to label 4 test tubes as follows: dH2O, Albumin, UNKNOWN, and
Milk. Put the test tubes in the well plate.
3. Use the Albumin pipet to measure 1 mL of the Egg white albumin stock solution
into the test tube named Albumin.
4. Use the dH2O pipet to measure 1 mL of distilled water into the corresponding
test tube.
5. Use the Milk pipet to transfer 1 mL of milk into the corresponding test
tube.
6. Add 24 drops of UNKNOWN from the bottle to the corresponding test tube.
7. Observe the color of the 4 test tubes and record.
8. Add 5 drops of Biurets reagent to each of the test tubes.
9. Observe and record the color.
10. Allow the samples to incubate for 5 minutes.
11. After 5 minutes, observe and record the color again.
12. A purple color means that is it a positive for protein. Record data.
13. Put aluminum foil on a flat surface and put 1 piece of filter paper on the
Aluminum foil.
14. Use a pipet to get the Sudan III.
15. Put 1 drop of Sudan III in 5 places on the filter paper.
16. Allow the drops to completely dry before going on to the next step.
17. Use a pencil to outline each of the 5 circles. Label the circles as: UNKNOWN,

Albumin, Distilled water, Milk, and Vegetable oil.

18. Use the dH2O pipet to put 1 drop of distilled water to right circle.
19. Use the Milk pipet to put 1 drop of milk to the milk circle.
20. Use the Albumin pipet to put 1 drop of albumin to the albumin circle.
21. Use the bottle of the UNKNOWN to add 1 drop of UNKNOWN to the corresponding
circle.
22. Use the bottle to put 1 drop of canola oil to the oil circle.
23. Add Sudan III to all circles. Allow it to sit for 30 minutes.
24. After 30 minutes. Observe and record whether the color leaked past the circle lines.
26. Wash and dry all equipment.
Results:
Benedicts Reagent
Sample
dH2O
Albumin
UNKNOWN
Milk
Glucose

Initial Color
Clear
Light yellow
clear
white
Milky white/clear

Final Color
blue
Blue and brown
Blue
purple and blue
Brown and blue

Sugar Present: y/n

N
Y
N
N
Y

Initial Color
clear
clear
Clear
White
Milky Clear
Yellow/brown

Final Color
Red/yellow
Clear/brown
Red/yellow
White/brown
Black
Black

Starch: Y/N
N
N
N
N
Y
Y

Lugols Iodine
Sample
dH2O
Albumin
UNKNOWN
Milk
Starch
Raw potato
Biurets Reagent
Sample

Initial Color

dH2O
Albumin
Unknown
Milk
Lipid

Blue/brown
Clear
Clear
White

Sample

Intermediate
Color
Light blue
Purple
Purple
Purple/white
Observations

Final Color

Protein: Y/N

Light blue
Purple
Purple
Purple/white

N
Y
Y
Y

Lipid: Y/N

Distilled water
Albumin
UNKNOWN
Milk
Vegetable Oil

Did not run outside lines

Stayed within circle
Stayed inside circle
Stayed inside circle
Went outside of lines

N
N
N
N
Y

Discussion/Conclusion:
In the test for reducing sugars, Albumin and Glucose tested positive. Both the Albumin
and glucose test tubes had brown, which is positive for sugar. The test that confused
me was the milk, because the milk has some sugar in it, so I figured it wouldve been
positive. I think the dH2O was the negative control because of common knowledge, we
know it does not contain sugar.
In the test for starch, the positive control was obviously starch. The negative control was
dH2O, because like I said before we know there isnt starch in it. I knew from prior
knowledge, the potato would be very black, and the test did in fact prove that. All others
were negative, and so the unknown is not a starch either.
In the test for protein, our negative control was the dH 2O once again, because we know
it is only water. Our positive control was the milk, because we know it contains protein,
and the test proved that when it turned purple. The Albumin and unknown also turned
purple, so we know the Unknown is a protein.
In the test for lipids, the vegetable oil was obviously the positive control in this test. The
distilled water was yet again our negative control. The only test that showed positive
was the vegetable oil because it went outside of the circle lines.
The unknown is a protein. I cannot identify what it is, so, I was incorrect. I can tell if it
was a lipid, protein, sugar, or starch, but not exactly what it is.
Works Cited
Campbell, Neil A. Biology: Concepts and Connections. San Francisco, CA: Pearson
Education, 2005. Print
Hands-on Labs, Inc.42-0291-00-02. HandsOn Learning. Lab Paq. Web. 02 Feb. 2016.
<http://www.HOLscience.com/>