Macromolecules Lab

Name: ___________________

Date: ____________________
Pre-Lab:
Watch the pre-lab video prepare a procedure in your lab notebook. Use the
internet or your textbook to answer the following questions on a separate
piece of paper (these do not have to be referenced).

1. What is the difference between a positive and negative control?

2. What is Benedicts Reagent?
3. What is a reducing sugar? (use a diagram)
4. What reaction occurs in a positive Benedicts Reagent test?
5. What causes the colour change in a positive Iodine test?
6. What is Biuret Reagent?
7. What causes the colour change in a positive Biuret test?
8. What reaction occurs in the emulsion test for fats?

Goal in the lab: Determine what macromolecule are in four unknown

powders/solutions using various biochemical tests. Confirm the results of the
tests by testing the known positive and negative controls as well.

Procedure

On the day of the lab:

The unknown substances will be in labelled containers at the front of the
class. One person from each pair will take one scoop of each of the unknown
substances into a labelled test tube (labelled either 1, 2, 3, or 4). Add 10 mL
of water to each test tube. These solutions will be used for the tests.
Complete predictions in the observations table.

A) Benedicts Test for Reducing Sugars

Rating Scale: Increasing amounts of reducing sugar

blue green orange red reddish brown

(-) (+) (++) (+++) (++++)

1. Add 2 mL of unknown solution into an empty, labelled test tube

2. Add 1.5 mL of Benedicts solution to the test tube
3. Mix gently. ** Make sure the test tube is labelled at the top!**
4. Place test tube in a boiling water bath
 5. Record the colour development in 3-5 minutes using the rating scale
above
6. Repeat with all 4 unknown solutions
7. Repeat with the controls

B) Starch Test for Polysaccharides

Amylose in polysaccharides is responsible for the formation of a deep blue

colour in the presence of iodine. The iodine molecule slips inside of the
amylose coil.

Brown (-) Blue-Black (++++)

1. Add 1 mL of unknown solution to an empty, labelled test tube

2. Add 1 drop of Iodine solution
3. Record colour development against a white paper placed behind the
test tube
4. Repeat with all 4 unknown solutions
5. Repeat with the controls

C) Biurets Test for Proteins

The Biuret reagent is made of sodium hydroxide and copper sulfate. The blue
reagent turns violet in the presence of proteins, and changes to pink when
combined with short-chain polypeptides.

1. Add 2 mL of unknown solution to an empty, labelled test tube

2. Add 4 drops of Biurets Reagent
3. Mix gently
4. Record colour development against a white background

Blue (-) Slight Purple/Pink (+) (polypeptide) Violet (++

++)(proteins)

D) Emulsion Test for Fats

1. Add unknown solution to an empty, labelled test tube

2. Add 2 mL of ethanol
3. Mex gently
4. Add 2 mL of deionized water
5. Observe any changes
Observations

Unknown/Food Benedicts Test Starch Test Biurets Test Fat Test

Solution Tested
Predict Actual Predict Actual Predict Actual Predict Actual

Unknown 1

Unknown 2

Unknown 3

Unknown 4

Carbohydrate test:

Positive Control
Monosachharide

Positive Control
Monosachharide
Positive Control
Polysachcharide

Negative Control
Polysachcharide

Protein test:

Positive Control
Protein

Negative Control
Protein

Lipid Test

Positive Control
Lipid

Negative Control
Lipid

Repeats if any: