Experiment No.

3
Lipids

Lipids are water – soluble organic compounds that can be extracted from cell and
tissues by non-polar solvents like ether, chloroform, benzene, etc. Lipids that contain
fatty acid are saponifiable, while those without fatty acid like steroids, are
non-saponifiable.

1. Physical Properties
A. Solubility Test
Reagents: chloroform, alcohol, 5% NaOH, 5%HCI, ether, coconut oil, oleic acid,
butter, margarine, lanolin, beef allow.

Procedure:
1. Place 1 ml. of each of the following in a clean dry test tube.
a. Chloroform
b. Alcohol
c. Water
d. NaOH
e. HCI
f. ether
2. To each test tube, add two drops of coconut oil and shake well. Observe.
3. Repeat the procedure using oleic acid, butter, margarine, lanolin, beef tallow.
4. Write your results in tabulated form.

B. Other Physical Properties:

Reagents: coconut oil, margarine, oleic acid, lanolin, beef tallow.

Procedure:
1. Observe and record the appearance, consistency, color and odor of each of the lipid
samples.
2. Write your answers in tabulated form.

C. Spotting Effect:
Reagents: alcohol, chloroform, coconut oil

Procedure:
1. Place a drop of coconut oil in a piece of filter paper.
2. On the same spot, place a drop of alcohol. Observe.
3. Repeat the procedure using chloroform.

Result:

a. oil and alcohol

b. oil and chloroform

3. CHEMICAL PROPERTIES:
A. Emulsification of Fats and Oils:
Reagents: coconut oil, butter, margarine, oleic acid, lanolin, beef tallow.

Procedure:
1. Place 3 drops of coconut oil into two dry test tubes.
2. Add 3 ml of hot water to each test tube.
3. Into the first test tube add 1 ml of soap solution.
4. Shake each test tube vigorously, then let stand and observe.

Result:
Oleic Acid: Test Tube 1

Test Tube 2

Coconut oil: Test Tube 1

Test Tube 2

Butter: Test Tube 1

Test Tube 2

Margarine: Test Tube 1

Test Tube 2

Cooking Oil: Test Tube 1

Test Tube 2

lanolin: Test Tube 1

Test Tube 2

B. Test for Unsaturation:

Reagents: oleic acid, butter/margarine, coconut oil, lanolin, beef tallow, CCI4,
bromine solution.
Procedure:
1. Test the substance listed below to determine the number of drops of Br2 solution (w/
CCI4 as the solvent) that can be decolorized. The first faint persistence of the color
bromine is the end-point of the reaction. Dissolve approximately 1 ml of each
substance to be tested in 5 ml of CCI4 before adding the bromine solution.
2. Enter the name of the substance tested as well as the amount (drops of bromine
solution) required.

Substance Tested Drops of Bromine solution absorbed

Oleic Acid
Butter/Margarine
Lanolin
Coconut Oil

C. Acrolein Test:
Reagents: KHSO4 crystals, coconut oil, lanolin, butter, margarine, beef tallow.

Procedure:
1. Place about 1 gram of KHSO4 in a dry test tube.
2. Add 1 ml of glycerol on the crystals of KHSO4.
3. Heat slowly at first to prevent evolution of SO2.
4. Then heat vigorously and note the odor of acrolein.
5. Repeat the procedure using the other lipid sample and compare the result with
glycerol.
6. Write your result in tabulated form.

D. Hydrolysis of Fat by Dilute Acids:

Reagents: 10% HCI, coconut oil, lanolin, oleic acid, butter, beef tallow.

Procedure:
1. Place 1 ml of coconut oil in a test tube.
2. Add 2 ml of 10%HCI.
3. Place in a water bath for 15 minutes.
4. Pour contents in an evaporating dish and allow to cool at room temperature.
5. Test with blue litmus paper.
6. Identify the fatty acid present.
7. Repeat the procedure using the other lipid samples.
8. Record your observations below.

Result:

E. Test for Cholesterol:

Reagents: fatty acids extract from Procedure D, chloroform, conc. sulfuric acid.

Procedure:
1. Place a few crystal of cholesterol in a clean dry test tube.
2. Add 2 ml. of chloroform and 5 drops of concentrated sulfuric acid.
3. Observe for the change in color of the solution from bluish red to purple.
4. Repeat the procedure using the fatty acid extract from procedure D.
5. Observe.