Kristine Cassandra S.

Ribleza
BSMLS 2G

EXPERIMENT 6
LIPIDS

POST-LAB GUIDE QUESTIONS

1. What is the purpose of translucent spot test? What is detected by this test? Is it
conclusive? Why?
- A translucent spot test is a preliminary test for lipids or fats in a given sample that
involves rubbing filter paper between the folds. Translucent spots indicate the
presence of fats, which are characterized by their translucence and greasiness.
Due to their fatty nature, the lipids will form a greasy or transparent spot that is
absorbed by the filter paper. Unlike lipids, a water spot sample on the paper will
evaporate. Fats are nonvolatile, meaning they have a very high boiling point and
do not evaporate easily, which is why they produce translucent spots. Unlike
water, lipids absorbed by paper cannot attain sufficient heat to evaporate at room
temperature. Therefore, a translucent spot remains visible when the filter paper is
held up to the light after the spot has dried.

2. Explain why results in the Test for Unsaturation are positive or negative based on the
structure of the lipid samples.
- Iodine is added to determine if a lipid is saturated or unsaturated. If the iodine
turns from brown to clear, the lipid is unsaturated, or the test for unsaturation was
positive. The iodine will bind itself to one of the double bonds, causing it to lose
its color. In unsaturated fats, iodine can integrate into the weaker double or triple
bonds, thereby losing its color and causing the fats to become clear. Indicating
the degree of lipid unsaturation, the amount of iodine required to achieve
decolorization is proportional to the number of double bonds in the lipid.
- If the iodine does not change color, it indicates that the lipid is saturated or that
the test for unsaturation is negative. Saturated fats turn brown and do not react
with iodine because they are composed of multiple single bonds, which are
 strong bonds. Since there is no space for an atom such as iodine to integrate
between carbon atoms and lose its color, saturated fats turn brown.
- Animal fats are often saturated in fatty acids, but vegetable oils are typically
unsaturated. A few plant foods, such as coconut oil, also contain a significant
amount of saturated fat. The composition of coconut oil is approximately 90%
saturated fats and 9% unsaturated fats. On the contrary, olive oil is composed of
86% unsaturated fats. The fatty acid components of lecithin can vary based on
the number of carbon atoms they contain and whether or not they are saturated
or unsaturated. The composition of fatty acids within a lecithin molecule
considerably influences its function. For instance, a lecithin molecule in which
both fatty acids are saturated facilitates oxygen intake in the lungs. Another
"species" of lecithin, containing two unsaturated fatty acids, is involved in
cholesterol transport in the blood. Usually, triglycerides generated from glycerol
and unsaturated fatty acids are oils. Those composed of saturated acids are
typically fats.

3. What is the principle behind Acrolein test? Explain why results showed positive or
negative.
- The Acrolein test is used to detect the presence of glycerol and fat. This test is
based on the dehydration reaction, in which the water molecules are removed
from the glycerol by adding the reagent potassium bisulfate (KHSO4). By heating
a fat sample in the presence of potassium bisulfate (KHSO4), which serves as a
dehydrating agent, acrolein (C3H4O, or CH2=CH-CHO) is formed and can be
easily detected by its odor. When fat is heated in the presence of a dehydrating
agent, the glycerol part of the molecule dehydrates to produce acrolein, an
unsaturated aldehyde with a pungent, irritating odor. The presence of fat or oil or
positive test is indicated by the pungent, irritating odor of acrolein. Whereas, the
absence of the pungent, irritating odor indicates a negative test for acrolein.

4. Explain the solubility test by comparing the structures of the lipid samples and the
solvents.
- The solubility test determines if a lipid is miscible or immiscible in polar and
non-polar solvents by comparing its solubility in different solvents. Thus, it is
based on the solubility of lipids in various solvents. Lipids are soluble in non-polar
 solvents such as chloroform, partially soluble in polar solvents such as ethanol,
but insoluble in polar solvents such as water. Therefore, if the sample produces
an oily layer above the water's surface, fat is present.
- Olive oil and coconut oil are insoluble in water due to the fact that their
molecules are nonpolar and water molecules are polar. In nature, oils are
hydrophobic, or “water fearing.” Since oil molecules are repelled by water, they
do not mix. Oil molecules will constantly float on top of water, forming an oil layer
on the surface. On the other hand, they are both soluble in cyclohexane due to
the fact that they are all nonpolar organic solvents. Olive and coconut oils are
insoluble in dilute HCl, an acid containing polar molecules, and thus do not
dissolve oil. In addition, they are insoluble in dilute alkali (dilute NaOH), which
forms a white colloidal substance when added.
- Lecithin is an amphiphilic compound with hydrophilic and water-soluble head
groups and hydrophobic and fat-soluble nonpolar tail groups. Partially soluble in
water, but rapidly forms emulsions when hydrated. The phosphatides that do not
include oil are soluble in fatty acids, but practically insoluble in fixed oils. Lecithin
is partially soluble in alcohol and almost insoluble in acetone when all
phosphatide portions are present.
- Due to the polyol groups' capacity to establish hydrogen bonds with water
molecules, glycerol is soluble in water. Glycerol has a specific gravity of 1.26,
which is slightly denser than water. This indicates that when glycerol is added to
water, it will sink to the bottom. It is only slightly soluble in organic solvents such
as ethyl acetate and diethyl ether, and it is insoluble in hydrocarbons.