Analysis

Lipids are organic compounds that are insoluble (non-polar) but soluble in organic
solvents and are fatty acids. Lipids are extracted from plants and animals and have mainly
hydrocarbons in their composition which are highly reduced forms of carbon. In this experiment,
we are able to perform experiments of lipids through Acrolein test, test for unsaturation, test for
phosphate, emulsification test, Lieberman-Burchard or acetic anhydride reaction, Carr-Price
reaction, and modified Furter-Meyer test.

The first test (Acrolein test) is used to detect the

presence of fat or glycerol. This is a qualitative test in
which the presence of a dehydrating agent (KHSO4)
results in the dehydration to form unsaturated aldehyde
which is acrolein with it’s characteristic odour. Our test
samples are glycerol, coconut oil, lecithin, and oleic acid
which are put in small test tubes and heated gently.
Glycerol and lecithin have positive results with a strong
acrid/pungent odor while the negative results; coconut
oil and oleic acid have oil-like smell only. This test tells
that glycerol and lecithin is positive in the presence of
fat/glycerol.

The second test(Test for unsaturation) is used to

detect the presence of double bonds in lipids. The
positive test in this is the change in color in which is the
same as the Hubl’s solution; red orange color. Our test
samples in this experiment are olive oil, oleic acid,
stearic oil, and coconut oil. 1 mL CHCl3 was added in
each test sample. We then added Hubl’s solution drop
by drop. Olive oil’s recorded drops were 194 drops, 108
drops for oleic acid, 6 drops for stearic acid, and 8 drops
for coconut oil. The results show that olive oil is the
most unsaturated test sample followed by oleic acid,
coconut oil, and stearic acid as the least saturated. It can
be seen in their structure that they contain double bonds (alkanes) which make them unsaturated.
 The third test (Test for Phosphate) is a test for
phospholipids. Lecithin is our test sample and a
positive test in this would be the formation of yellow
precipitate. It was first put in the porcelain crucible,
added with H2O, and was filtered. The filtrate was
then added 1 mL 5% (NH4) MoO4 and 3 drops conc.
HNO3. It was heated to boil, then few yellow
precipitate was present which still indicates that
lecithin has the presence of phosphate.

The fourth test (Emulsification

test) tests if solution can form an
emulsion with other substances. Our
test samples in this are; (1) 1 mL
coconut oil + 5 drops .1% bile salt
solution, (2) 1 mL coconut oil + 5
drops H2O + tiny crystals of
cholesterol (3) 1 mL olive oil + 5 drops 1% aqueous solution of lecithin. All the test samples are
put in a test tube and are mixed very well. The first and third test sample showed a positive result
while the second did not. The first and third test sample showed a positive result in which the
liquids mixed in them became miscible was because they contain an emulsifying reagent while
the second test sample does not.

The fifth test ( Lieberman-Burchard or Acetic

Anhydride reaction) is a test for cholesterols. Our test
sample in this are cholesterol and bile salts. A positive
test would be a change in color. It was placed in a test
tube with 1 mL CHCl3. 5 drops of acetic anhydride and a
drop of conc. H2SO4 was added. It was then mixed very
well and the color was observed. The positive result in
this test is the first sample which is cholesterol. The
result in this is a change in color which is because of the
hydroxyl group of cholesterol which reacts to the reagent
( acetic anhydride) and conc. H2SO4 and also the
increasing conjugation of a substance to a hydrophilic state.
 The sixth test ( Carr-Price test) is used to
detect presence of vitamin A and related carotinoids.
We put 1 mL of chloroform in a test tube and added 2
drops of cod liver oil. After that it was cooled and
added 2 mL of cold saturated chloroform solution
SbCl3. Changes in color were observed and it was
tested again with coconut oil and olive oil. A positive
result would be the change in color. In our result the
first sample gave a positive result which is cod liver
oil. Cod liver oil contains very high levels of vitamin D
and vitamin A that’s why its result is positive.

The last test (Modified Furter-

Meyer test) is used to detect the
presence of tocopherols by giving a
bronze-red solution. We put 1 mL
.05% alpha-tocopherol in CHCl3 in a
test tube which was added 3.5 mL n-
butyl alcohol and .5 mL conc HNO3.
We did all the procedure also on a
commercially available vitamin E
(Myra-E). We then mixed and placed it in water bath and noted its color. Only α-tocopherol is
recognized to meet human requirements.

Conclusion

The purpose of Experiment 8 is to test for lipids using 7 tests and these tests are Acrolein
test, Test for Unsaturation, Test for Phosphate, Emulsification test, Lieberman-Burchard Test or
the Acetic Anhydride Reaction, Carr-Price Test, and Modified Furter-Meyer test. First, Acrolein
test, we had 4 test samples (glycerol, coconut oil, lecithin, and oleic acid) and among the 4 test
samples, glycerol and lecithin are the test samples which tested positive for fats. Second, Test for
Unsaturation, to detect presence of double bonds in lipids has 4 test samples namely oleic acid,
stearic oil, coconut oil and olive oil and among these test samples olive oil showed that it is the
most unsaturated having 194 drops of Hubl’s solution and stearic acid as the least unsaturated
with only 6 drops. Third, Test for Phosphate has 1 test substance (Lecithin) which showed signs
of a yellow precipate and is an indication for the presence of phosphate. Fourth, Emulsification
test, to test if possible to form emulsion with other substances, it has 3 test solutions (1 ml
coconut oil + 5 drops of 1% bile salt solution, 1 ml coconut oil + 5 drops H20 + tiny crystals of
cholesterol and 1 ml olive oil + 5 drops of 1% aqueous solutions) only the first and third test
solutions showed a positive result. Fifth, Lieberman- Burchard or Acetic Anyhydride, test
samples are cholesterol and bile salts, the positive result in this experiment is the first sample;
cholesterol, which has a change in color. Sixth, Carr-Price Test, to detect presence of Vitamin A,
Test samples are Cod liver oil, coconut oil and olive oil. Cod had a positive result having a
change in color. Last, Modified Furter-Meyer test, to detect presence of Tocopherols, test
samples are vitamin E and 0.5 % alpha-tocopherol. Light brown color appeared in alpha-
tocopherol which indicated presence of tocopherols.