Name:Click or tap here to enter text.

Laboratory 2: Biological Molecules (“McMush Lab”)

Cells are made up of small molecules like water, ions such as sodium and magnesium, and large
carbon-based molecules. There are four important types of large carbon-based molecules
(Biological Molecules/Organic Macromolecules) in living organisms- proteins, carbohydrates
(sugars & starches), lipids (fats), and nucleic acids. Proteins, carbohydrates, and fats serve as
nutrients in the food that we eat. In this lab, we will be investigating three of the four important
Biological Molecules: Carbohydrates (sugar/starch), Proteins, and Lipids (fats).
Carbohydrates are used by living organisms as an important source of energy. The monomers
(building blocks) of carbohydrates are referred to as monosaccharides. Common examples of
monosaccharides include glucose, fructose, galactose, ribose, and deoxyribose. Sucrose, or table
sugar, and lactose, the sugar found in milk, are double sugars (disaccharides) made from two
monosaccharides. Benedict’s Solution can be used to test for the presence of a monosaccharide,
like glucose. In the presence of glucose, Benedict’s Solution turns from blue to orange when
heated (orange is positive, blue is negative).
When many monosaccharides join together, the resulting molecule is called a polysaccharide.
Important polysaccharides include cellulose, starch, and chitin. Lugol’s Iodine can be used to test
for the presence of a polysaccharide, like starch. In the presence of starch, Lugol’s Iodine turns
from amber to dark blue (dark blue/black is positive, amber is negative).
Lipids are organic compounds that are do not dissolve in water, examples include fats, oils, and
the wax covering leaves. The large number of carbon to hydrogen bonds serves to make lipids
energy-rich storage molecules. Most dietary fats that we consume are called triglycerides which
are composed of a glycerol and three fatty acids. Sudan IV Solution can be used to test for the
presence of a lipid. In the presence of a lipid-rich solution, Sudan IV turns a bright red color
(bright red is positive, dark red is negative).
Proteins are made of monomers (building blocks) called amino acids, which are composed of
atoms of carbon, hydrogen, oxygen, and nitrogen. Proteins are large and complex molecules that
combine to form various components of living organisms such as muscle fibers, enzymes, and
hemoglobin. Proteins are made from specific sequences of amino acids. Biuret’s Solution
(Cu2SO4 + NaOH) can be used to test for the presence of protein. In the presence of protein,
Biuret’s Solution turns from blue to violet (violet is positive, blue is negative).

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Table 1: Indicators Used in this Lab
TEST BIOMOLECULE NEGATIVE POSITIVE
REAGENT/INDICATOR TESTED FOR RESULT RESULT
Lugol’s Iodine (LI) Carbohydrate (Starch) Yellow/orange Blue/Black Color
Benedict’s Solution Carbohydrate Blue Green, Red,
(Sugar/glucose) Orange, Yellow
Biuret’s Solution Protein Blue Violet
Sudan IV solution Lipids/Fats Brown Bright Red

You will be using these indicators to not only determine whether the Biomolecule is present
(positive control), but also if the biomolecule is NOT present (negative control). Each
experiment you perform will have a negative control and a positive control as baseline data to
compare to. Finally, you will be given an “unknown” sample in which you will perform all of
the above tests and using your knowledge of negative and positive controls, determine whether
you have the biomolecule or not.
Watch these videos to see how the indicators work!
1. Watch this first video to demonstrate the color change with Iodine, Biuret and Benedicts
(watch up until 3:06 mins): https://www.youtube.com/watch?v=sLP8dcnWnJg

2. Watch this video to see demonstrate the color change with Sudan IV. Note how the
Sudan IV turns bright red in the presence of lipids. https://www.youtube.com/watch?
v=pn8ZN3QAVtE

Objectives
1. Students will be familiar with the names of the indicator solutions above.
2. Students will be able to interpret negative and positive results from the indicator solutions
above.
3. Students will use the above indicator solutions to test for the presence or absence of biological
macromolecules found in an unknown food sample.

Procedure
For the following Biomolecule tests, each group will receive an unknown food slurry/sample.
You will be using this same unknown in all four tests today. You will be asked to hypothesize
what biomolecules might be found in a blended Happy Meal® slurry (AKA McMush!). Your
unknown may or may NOT have all the Biomolecules we are testing for as you complete all four
experiments.

For each experiment:

 Test Tube 1 will always contain distilled water as the negative control
 Test Tube 2 will contain either glucose, starch, albumin, or vegetable oil as a
positive control
 Test Tube 3 will contain the “McMush Slurry” as the experimental unknown.

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Laboratory 2: Biological Molecules (“McMush Lab”) Pre-
Laboratory Assignment
Please complete the tables/questions below after reading Laboratory 2. You may also want to review the
topic of Biological Molecules in Chapter 2 of your textbook.
1. During our investigation of the following Biomolecules: Lipids, Proteins,
and Carbohydrates (glucose, starch), we will be using several Test Reagent/Indicators that will test for
each of these. Using the information found in Laboratory 2, fill in the table below:

Macromolecule Test Reagent Positive Test Negative Test

Color Color
 Click or tap here  Click or tap here to  Click or tap here to
Carbohydrate (glucose) to enter text. enter text. enter text.

 
 Click or tap here  Click or tap here to  Click or tap here to
Carbohydrate (starch) to enter text. enter text. enter text.

 
 Click or tap here  Click or tap here to  Click or tap here to
Proteins to enter text. enter text. enter text.

 
 Click or tap here  Click or tap here to  Click or tap here to
Lipids to enter text. enter text. enter text.

2. Define the following:

a. Negative Control: Click or tap here to enter text.

b. Positive Control: Click or tap here to enter text.

3. Thinking about the procedure for Lab 2, which test tube # is considered the negative control?

Click or tap here to enter text.

4. Thinking about the procedure for Lab 2, which test tube # is considered the positive control?

Click or tap here to enter text.

5. Why do we need to use the Test Reagents (that you indicated in the above table)?

Click or tap here to enter text.

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6. Finally, we will be investigating what Biomolecules might be found in a “McMush Food Slurry”.
What biomolecule (s) do you think would be found in a typical Happy Meal® from McDonalds®? In
your answer, be sure to list what food in the Happy Meal® would contain that particular Biomolecule.
Assume the meal contains, a cheeseburger, french fries, and a Sprite® soft drink.

Click or tap here to enter text.

1. Testing for Carbohydrates (Sugar/Glucose)

1. Three test tubes were filled with the following:

a. Test tube 1 to the 1 cm mark with distilled water and Benedicts’ reagent to the 3 cm
mark.
b. Test tube 2 to the 1 cm mark with glucose solution and Benedicts’ reagent to the 3
cm mark.
c. Test tube 3 to the 1 cm mark with your UNKNOWN McMush Slurry and
Benedicts’ reagent to the 3 cm mark.
2. Contents of each tube were mixed thoroughly by holding the tube between the thumb and
index finger of one hand and gently thumping the tube.
3. Benedicts indicator needs to be boiled to be activated. Tongs were used to place the test
tubes in a hot water bath for 5 minutes.  The results are shown below in Figure 1.
Record the color in the data table below (Table 2).

Figure 1: Results for the Benedicts Test for sugar. Tube 1 is blue, Tube 2 is orange, Tube 3 is
blue.

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Table 2: Carbohydrate (Sugar/Glucose) Results

Test Tube # Test Tube 1 (d-H2O) Test Tube 2 Test Tube 3

(glucose) (Unknown)
Blue Orange Click or tap here to
Color Change After enter text.
Boiling

*Note: Benedicts’ Reagent is initially sky blue. Refer to Table 1 at the beginning of the lab for a reminder of negative and positive results
with this reagent.

Conclusions: Be sure to explain why a particular tube changed color or didn’t change
color. Don’t just indicate that a tube changed or didn’t change color, explain WHY!

Test Tube #1: Test tube 1 is blue. This indicates a negative result. Sugar is NOT present in
this tube.

Test Tube #2: Test tube 2 changed from blue to orange. This is a positive result. Sugar is
present.

Test Tube #3: Click or tap here to enter text.

2. Testing for Carbohydrates (Starch)

1. Three test tubes were filled with the following:

a. Test tube 1 to the 1 cm mark with distilled water and Lugol’s Iodine reagent
to the 3 cm mark.
b. Test tube 2 to the 1 cm mark with starch solution and Lugol’s Iodine reagent
to the 3 cm mark.
c. Test tube 3 to the 1 cm mark with your UNKNOWN McMush Slurry and
Lugol’s Iodine reagent to the 3 cm mark.
2. Contents of each tube were mixed thoroughly by holding the tube between the thumb and
index finger of one hand and gently thumping the tube.
3. The results are shown below in Figure 2. Record the color in the data table below (Table
3).

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 Figure 2 Lugol's Iodine test for starch results. Test tube 1 is orange, test tube 2 is blue/black,
and test tube 3 is blue/black.

Table 3: Carbohydrate (Starch) Results

Depression Well # #1 (d-H2O) # 2 (Starch) # 3 (Unknown)

Click or tap here to Click or tap here to Click or tap here to
Color Change After enter text. enter text. enter text.
Stirring

*Note: Lugol’s is initially amber. Refer to Table 1 at the beginning of the lab for a reminder of negative and positive results with this
reagent.

Conclusions: Be sure to explain why a particular well changed color or didn’t change
color. Don’t just indicate that a particular well changed or didn’t change color, explain
WHY!

Test tube #1: Click or tap here to enter text.

Test tube #2: Click or tap here to enter text.

Test tube #3: Click or tap here to enter text.

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3. Testing for Proteins

1. Three test tubes were filled with the following:

a. Test tube 1 to the 1 cm mark with distilled water and Biuret reagent to the
3 cm mark.
b. Test tube 2 to the 1 cm mark with albumin and Biuret Iodine reagent to the 3 cm
mark.
c. Test tube 3 to the 1 cm mark with your UNKNOWN McMush Slurry and Biuret
reagent to the 3 cm mark.
2. Contents of each tube were mixed thoroughly by holding the tube between the thumb and
index finger of one hand and gently thumping the tube.
3. The results are shown below in Figure 3. Record the color in the data table below (Table 4).

Figure 3 Biuret test for proteins results. Test tube 1 is blue, test tube 2 is purple, and test tube 3
is purple.

Table 4: Protein Results

Depression Well # #4 (d-H2O) # 5 (Albumin) # 6 (Unknown)

Click or tap here to Click or tap here to Click or tap here to
Color Change After enter text. enter text. enter text.
Stirring

*Note: Biuret is initially blue. Refer to Table 1 at the beginning of the lab for a reminder of negative and positive results with this reagent.

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Conclusions: Be sure to explain why a particular well changed color or didn’t change
color. Don’t just indicate that a well changed or didn’t change color, explain WHY.

Test tube #1: Click or tap here to enter text.

Test tube #2: Click or tap here to enter text.

Test tube #3: Click or tap here to enter text.

4. Testing for Lipids

1. Three test tubes were filled with the following:

a. Test tube 1 to the 1 cm mark with distilled water and 3 drops of Sudan IV reagent.
b. Test tube 2 to the 1 cm mark with vegetable oil and 3 drops of Sudan IV reagent.
c. Test tube 3 to the 1 cm mark with your UNKNOWN McMush Slurry and 3 drops of
Sudan IV reagent.
2. Contents of each tube were mixed thoroughly by holding the tube between the thumb and
index finger of one hand and gently thumping the tube.
3. The results are shown below in Figure 4. Record the color in the data table below (Table 5).

Figure 4 Sudan IV test for lipids results. Test tube 1 is a brown powder, test tube 2 is dissolved
bright red, and test tube 3 is dissolved bright red.

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Table 5: Lipid Results

Depression Well # #7 (d-H2O) #8 (Vegetable Oil) #9 (Unknown)

Click or tap here to Click or tap here to Click or tap here to
Color Change After enter text. enter text. enter text.
Stirring

*Note: Sudan IV is initially brown solution. Sudan IV will turn a bright red color and become somewhat thicker for a positive Lipid test.

Conclusions: Be sure to explain why a particular well changed color or didn’t change
color. Don’t just indicate that a tube changed or didn’t change color, explain WHY!

Test tube #1: Click or tap here to enter text.

Test tube #2: Click or tap here to enter text.

Test tube #3: Click or tap here to enter text.

Summarize your results of the “Unknown McMush Slurry” in test tube 3:

Table 6: UNKNOWN McMush Slurry Results

+ = Biomolecule is present; - = Biomolecule is NOT present
Unknown Carb (Sugar) Carb (Starch) Protein Lipid
Sample
Click or tap here Click or tap here Click or tap here Click or tap here
McMush Slurry to enter text. to enter text. to enter text. to enter text.

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Conclusions

1. All of the experiments we did today had distilled water in Test tube #1. Explain the
significance of the distilled water. Why did we use it?

Click or tap here to enter text.

2. Look back at your results from each of the 4 experiments. Did test tube #2 show a positive or
negative result for the Biomolecule tested? Explain! Hint: What was the significance of these
tubes?

Click or tap here to enter text.

3. In the pre-lab, you were asked to hypothesize what Biomolecules might be present in a
blended Happy Meal® (McMush Slurry). If you still aren’t sure, go to
https://www.mcdonalds.com/ and do some research on what Biomolecules should be present
(assume the Happy Meal® contains a cheeseburger, French fries, and a Sprite® soft drink).
Now that you have that information, did your UNKNOWN sample contain all the Biomolecules
that SHOULD be present in a happy meal? If not, which molecule(s) were missing? What
SHOULD be present in a McMush Slurry. What evidence can you provide?

Click or tap here to enter text.

LICENSES AND ATTRIBUTIONS

 This work is a derivative of the McMush Lab. Authored by: Shannon Muskopf. Located at:
https://www.biologycorner.com/worksheets/mcmush.html. License: Creative Commons
Attribution-NonCommercial-ShareAlike 4.0 International License.
 Lab 2 Biological Molecules “McMush Lab” was created by Michele Finn using the above
resource and making several changes, additions, and deletions.

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