LIPIDS

Procedure:
Test for unsaturation
1. Shake 1ml of oil sample with 5ml of ether (benzene) and add 3 drops of chlorine.
Observations:
The oil sample mixed with the ether (benzene) because both compounds are nonpolar. This follows the thumb rule like
dissolves like.
2. Shake again and note the changes of chlorine in water.
Observations:
Even after shaking, chlorine still did not mix with the oil+benzene solution. The large chlorine droplets diffused into
tiny droplets, though. This is due to shaking.
What do the results in 1 & 2 indicate?
Chlorine is usually added to oil/ together with an organic compound for the process termed as halogenation. Chlorine
molecules are supposed to attach themselves to the double bonds of the fatty acids. Since chlorine droplets did not mix
with the solution, this means that the oil this time was saturated or perhaps, the unsaturated carbons of the oil are now
filled with the chlorine molecules and what is now observable are the excess chlorine molecules.

Saponification
1. In a test tube, put 2 drops of oil sample and add 6ml of 20% NaOH and mix. Keep in a boiling water bath for 20mins,
mix occasionally and let it cool.
Observations:
After heating, A cloudy appearance was noticed in the solution. Also, formation of precipitates were noted.
2. Divide the clear solution into three test tubes.
To the first test tube, add concentrated HCl drop by drop with mixing: note the scum forms at the top.
Observations:
In the experiment, a layer of small bubbles was formed after adding HCl. Smaller bubbles indicated that HCl was
somehow dissolved into smaller particles . Moreover, oil has a lower specific gravity than HCl , thus floats on water.
What indicates the scum formation?
If the pH of a soap solution is lowered by acidic contaminants, insoluble fatty acids precipitate and form a scum. A
second problem is caused by the presence of calcium and magnesium salts in the water supply (hard water). These
divalent cations cause aggregation of the micelles, which then deposit as a dirty scum.
Second test tube: add 2ml of NaCl solution. Note the precipitate formed.
What is the color of the precipitate?
Observations:
A white precipitate was formed on the surface after NaCl solution was added. The white precipitate formed is the soap
which is water insoluble.
Third test tube: Add 10 ml of water, shake vigorously and add 2ml of phenolthalein. Observe the change in color.
Observations:
After adding phenolphthalein to the solution, the upper portion changed in color, from white to magenta/pink whereas
the lower portion of the solution remained the same.

Phenolphthalein is used to test for the presence of free fatty acids as well as the pH of the solution. Pink coloration
indicates that the pH of a solution is 8.3-9. When the pH drops below pH 8.3 phenolphthalein goes colourless. One
example is an alkaline solution of milk. Together with lipase and phenolphthalein, the solution will change from pink
to colourless as the fat in milk is broken down to form fatty acids (and glycerol) thus reducing the pH to below 8.3.

Answer briefly:
1. Compare the properties of lipids with those of carbohydrates.
Carbohydrates are hydrophilic. The presence of hydroxyl groups allow the carbohydrates to interact with the aqueous
environment and to participate in hydrogen bonding both between and within chains. In contrast, lipids are known for
their water insoluble property, that is, it is repulsive from water molecules but are soluble in nonpolar compounds.
In terms of energy production, via oxidation, a gram of carbohydrate can produce about 4.5kcal while lipids can
produce more than twice as much (9kcal). Carbohydrates can form a polymer while lipids cannot.
2. What characteristics do all lipids have in common?
Water insoluble. Mostly are composed of fatty acids.
3. What is the difference between a fat and oil? Between a fat and wax?
Fats
- solid at room temp
- from animal sources, hence, animal fats
- contain a greater proportion of saturated fatty acids
Oils
- liquid at room temp
- from plants, hence, vegetable oils
- have greater concentration of unsaturated fatty acids
Wax

- an ester of a saturated fatty acid and a long- chain alcohol, each containing from 14 to 30
carbon atoms

4. What are the three major types of lipids found in cell membrane?
Phospholipids

- major component of the cell membrane; forms the lipid bilayer sheet which is the primary barrier
between the internal and external cellular environment.

Glycolipids - The heads of glycolipids (glyco- stands for sugar) contain a sphingosine with one or several sugar
units attached to it. The hydrophobic chains belong either to the two fatty acids (FA) or one FA and the hydrocarbon
tail of sphingosine - in the case of sphingomyelin and the glycolipids.
Cholesterol - membrane fluidity regulator promoting rigidity of the cellular membrane

5. Describe the test for the presence of fat or oil.

EXPERIMENT
SOLUBILITY TEST
Food sample + water
Food sample + chloroform(CHCl3)

OBSERVATION

INFERENCE

Does not dissolve

Oil / fat present.

SPOT TEST
Smear the food sample on paper.
ACROLEIN TEST
Food sample + KHSO4. Heat

A translucent spot is
observed.

Oil / fat present.

An irritating odour is
obtained.

Oil / fat present.