I.

Theory

Lipids are a heterogenous group of compounds, including fats, oils,

steroids, waxes, and related compounds, which are related more by their
physical than by their chemical properties. The lipids are a class of
compounds distinguished by their insolubility in water and solubility in
nonpolar solvents. Lipids are important in biological systems because they
form the cell membrane, a mechanical barrier that divides a cell from the
external environment. Together with lipids, the carbohydrates and proteins,
are the primary components of plant and animal cells. Cholesterol and
triglycerides are lipids. The lipids are readily stored in the body, serve as a
source of fuel, and are essential components of cell structure. Lipids have
the common property of being relatively insoluble in water and soluble in
nonpolar solvents such as ether and chloroform. They are important dietary
constituents not only because of their high energy value but also because of
the fat-soluble vitamins and essential fatty acids contained in the fat of
natural foods. Lipids may be classified according to their hydrolysis products
and according to similarities in their molecular structures. On the other hand,
the different tests that will determine the presence of lipids is Solubility test
which detects lipid solubility in various solvents to check whether it is
miscible or immiscible in polar or non-polar solvents, Translucent spot test
a preliminary test for the lipids, which is characterized by a translucent and
greasy spot, Acrolein test which used to detect the presence of glycerol or fat
and the last test is Huble’s test is used to detect the degree of unsaturation
in oil or fat.

II. Different Test for Lipids

1. Solubility Test

The preliminary test detects the presence of all lipids. The lipid
solubility test determines whether the lipid is miscible or immiscible in
polar or non-polar solvents. As a result, it is based on the characteristic
of lipid solubility in various solvents. Lipids are easily miscible in non-
polar solvents such as chloroform, partly soluble in polar solvents such
as ethanol, and immiscible in polar solvents such as water.

Materials and Apparatus

1. Test tubes 7. Clarified butter

2. Test tube holder 8. Vegetable Oil
3. Bunsen burner 9. Linseed Oil
4. Dropper 10. Alcohol
5. Lab aprons 11. Chloroform
6. Hand Gloves 12. Water
 Procedure

1. Take three test tubes which contains 5ml of given sample solution
to be tested and label each test tubes A, B and C.
2. Then, add different solvents like water, ethanol, and chloroform in
each test tubes A, B and C.
3. Shake the tubes and allow it to stand for 1 minute.
4. Observe the change in solubility of the given sample.
5. If it is soluble in water then fat is absent, if its sparingly soluble in
ethanol then fat is present and same for chloroform.

2. Translucent Spot Test

A translucent spot test is a preliminary test for lipids that is

distinguished by a transparent and greasy area. Unlike water, the lipid
will not moisten the filter paper. Because of their greasy nature, the lipids
will permeate the filter paper and produce a greasy or transparent spot.
Unlike lipids, the water stain will vanish off the paper.

Materials and Apparatus

1. Filter paper 5. Lab aprons

2. Candle 6. Vegetable Oil
3. Dropper 7. Clarified butter
4. Hand gloves 8. Linseed Oil

Procedure

1. Take the sample to be tested, press a little in the folds of the filter
paper.
2. On folding if there is the appearance of greasy spot indicates the
presence of oils or fats.
3. The spot grows larger on heating and drying the filter paper.

3. Acrolein test

The acrolein test detects the presence of glycerol and fat. This test is
based on the dehydration process, which removes water molecules from
glycerol by using the reagent potassium hydrogen sulphate. The
interaction between glycerol and potassium hydrogen sulphate produces
acrolein, which is distinguished physically by the emission of a strong
odor.

Materials and Apparatus

1. Test tubes 6. Vegetable Oil

2. Test tube holder 7. Linseed Oil
3. Bunsen burner 8. Hand gloves
4. Potassium bisulfate 9. Lab aprons
 5. Clarified butter

Procedure

1. Take the sample to be tested in a test tube.

2. Add few crystals of potassium bisulfate to it.
3. Heat the mixture and observe the change in odor.
4. If there is pungent irritating odor then the presence of fate or oil is
confirmed.

4. Huble’s Test

This test determines the degree of unsaturation in a particular

sample. If the violet color of iodine fades after reaction with Huble's
reagent, the oil is unsaturated; if the color persists, the fat or oil is
saturated.

Materials and Apparatus

1. Test tube 5. Huble’s reagent

2. Test tube holder 6. Cotton seed oil
3. Dropper 7. Linseed Oil
4. Hand gloves 9. Chloroform
5. Lab aprons

Procedure

1. Take two test tube and add 3ml of chloroform in each.

2. Add cottonseed oil in one and linseed oil in the second test tube.
3. Shake the mixture well and add 3 drops of Huble’s reagent in
each test tube.
4. The violet color of iodine fades away in linseed oil test tube while
the color does not fade away in cottonseed oil.
5. This shows that linseed oil is more unsaturated than cotton seed
oil.

Conclusion

We therefore conclude that the following experiments that we watched

enabled us to determine the presence and characteristics, as well as the
chemical properties of lipids. The analyst became more knowledgeable
about the presence of lipids in the following test that were conducted. Lipids
are fatty, waxy, or oily compounds that are truly hydrophobic and insoluble in
polar solvents such as water, but soluble in organic solvents. Based on the
result of the tests, the following conclusion were made. For Solubility test,
clarified butter, vegetable oil and linseed oil are immiscible in water while in
ethyl alcohol and chloroform the samples are miscible. The fatty acids are
immiscible in water because they are composed mostly of hydrogen bonds
found in lipids are considered nonpolar. Following the solubility principle
which is like dissolve like, the sample is partially miscible in ethyl alcohol and
miscible in chloroform because both are nonpolar organic compounds.

For the Translucent spot test, all the three samples resulted positive
which means there is an appearance of translucent and greasy spot which is
evident in the filter paper. Most grease or fat have a high boiling point, so
they are non-volatile. The spot of grease can never absorb enough heat to
evaporate. When the liquid is inside the sheet of paper, it diffracts light. So,
light can pass from one side of the paper to another side which results to a
translucent effect. For the Acrolein Test, the three samples yielded a positive
result of burnt cooking oil smell or pungent irritating odor. This means that
these sample contain glycerol. For the Acrolein Test, the three samples
yielded a positive result of burnt cooking oil smell or pungent irritating odor.
This means that these sample contain glycerol. For the Huble’s Test, the
cotton seed oil resulted in violet color which does not fade away and in
linseed oil it resulted in violet color which fade away. This means that linseed
oil is more unsaturated than cotton seed oil. Contrarily, the different tests
performed in this experiment determined the chemical properties of lipids.