BIOCHEMISTRY LABORATORY FOR BS NURSING

NAME _______________________________________________ DATE ______________

COURSE & SECTION _________________ SEMESTER & YEAR ___________________

ACTIVITY NO. 5

QUALITATIVE ANALYSIS OF LIPIDS

i. Concept

The qualitative analysis of lipids helps us determine the presence or absence of

lipids, depending upon the color change. Lipids are organic biomolecules soluble in non-
polar solvents like chloroform, ether, acetone, etc., and insoluble in water.

In nature, different varieties of lipids show structural diversity among each other.
Some common lipids are fatty acids, soaps, fats, oils, waxes, phospholipids, etc. Here, we
will discuss the different qualitative methods for analyzing lipids.

A. Solubility Test - It is the preliminary test that detects the presence of all lipids.
Solubility test detects lipid solubility in various solvents to check whether it is miscible
or immiscible in polar or non-polar solvents. Thus, it is based on the property of lipid
solubility in different solvents. Lipids are readily miscible in non-polar solvents like
chloroform, partially soluble in a polar solvent like ethanol, and immiscible in a polar
solvent like water.

Materials: Reagents (Chloroform, Cold Alcohol)

Fats & Lipids samples (coconut oil, glycerol, any cooking oil)
Distilled water
Procedures:

1. Take the lipid sample in three different test tubes by labeling it as A, B, and C.
2. Add different solvents like water, ethanol, and chloroform in each test tube A, B
and C.
3. Shake the tubes and allow it to stand for 1 minute.
4. Check the solution for whether the lipid is soluble or insoluble.
5. Observe and record your observations and note which substances exhibited
solubility.

Answer the following:

1. What is the purpose of the solubility test in lipid analysis?

2. How does the solubility of lipids vary depending on their chemical structure?
3. What should be the criteria for determining whether a lipid is soluble or insoluble I
a particular solvent?
B. Translucent Spot Test - This is also a preliminary test for lipids, characterized by a
translucent and greasy spot. The lipid will not wet the filter paper, unlike water. Due to
their greasy texture, the lipids will form a greasy or translucent spot, and penetrate the
filter paper. Unlike lipids, the spot of water will disappear from the paper.

Procedure:

1. Take a filter paper.

2. Add one drop of water at one end and a drop of oil or lipid at the other end.
3. Observe the appearance of a translucent spot on the filter paper.

C. Emulsification Test – This is used to detect the presence of lipids. It is the process
that stabilizes the water and oil emulsion by using emulsifying agents. The lipid or oil
in water will appear as the supernatant. The high surface tension of water develops a
separate layer by adding emulsifying agents like bile salts, soap, etc. Emulsifying
agents emulsify the lipid, after which the lipids appear as tiny droplets suspended in
the solution.

Materials: Test substances (e.g., egg yolk, mayonnaise, coconut oil, water, non-
emulsifying substances like sugar solution). 5 Test tubes, Graduated
cylinder or pipette, Glass stirring rod, Water, Beakers or small cups,
Watch or timer

Procedure for the Preparation of Test Solutions:

1. Label test tubes with the names of the substances to be tested and add a small
amount (e.g., 2 mL) of each test substance into the corresponding test tube.
2. Add an equal volume of water (3 ml) to each test tube containing the test
substance.
3. Insert a glass stirring rod into each test tube and vigorously stir the mixture for a
set period (e.g., 1 minute).
4. Allow the mixtures to stand undisturbed for a specified period (e.g., 5 minutes) to
observe any separation or emulsification.
5. Observe and record whether the mixture remains stable (emulsified) or separates
into distinct layers.
6. Compare the behavior of the test substances. Note which substances show
emulsification and which do not.
7. Record your observations and note which substances exhibited emulsification.

Discussion:

Discuss the results with your peers. Consider possible explanations for any
unexpected outcomes. Explore the significance of emulsification in various
applications (e.g., food preparation, cosmetics, pharmaceuticals).

Conclusion:

Summarize the findings of the experiment. State which substances

demonstrated emulsification, indicating the presence of lipids. Reflect on the reliability
and limitations of the Emulsification Test.